WO2002066643A2

WO2002066643A2 - Proteins, polynucleotides encoding them and methods of using the same

Info

Publication number: WO2002066643A2
Application number: PCT/US2001/048732
Authority: WO
Inventors: Uriel M. Malyankar; Suresh G. Shenoy; Kimberly A. Spytek; Bryan D. Zerhusen; Meera Patturajan; Xiaojia Guo; Ramesh Kekuda; Esha A. Gangolli; Richard A. Shimkets; Raymond J. Taupier, Jr.; Li Li; Muralidhara Padigaru
Original assignee: Curagen Corporation
Priority date: 2000-11-13
Filing date: 2001-11-13
Publication date: 2002-08-29
Also published as: WO2002066643A3

Abstract

Disclosed herein are nucleic acid sequences that encode novel polypeptides. Also disclosed are polypeptides encoded by these nucleic acid sequences, and antibodies, which immunospecifically-bind to the polypeptide, as well as derivatives, variants, mutants, or fragments of the aforementioned polypeptide, polynucleotide, or antibody. The invention further discloses therapeutic, diagnostic and research methods for diagnosis, treatment, and prevention of disorders involving any one of these novel human nucleic acids and proteins.

Description

PROTEINS, POLYNUCLEOTIDES ENCODING THEM AND METHODS

O USING THE SAME

FIELD OF THE INVENTION

The invention relates to polynucleotides and the polypeptides encoded by such polynucleotides, as well as vectors, host cells, antibodies and recombinant methods for producing the polypeptides and polynucleotides, as well as methods for using the same.

BACKGROUND OF THE INVENTION

The present invention is based in part on nucleic acids encoding proteins that are new members ofthe following protein families: Membrane protein/neuropilin/metalloproteinase- like protein-like, Fibrillin-like, KIAAl 589-like, WD 40 motif-like, Opioid Bing Cell Adhesion Molecule-like, Triacylglycerol lipase-like, IGE Receptor Beta Subunit-like, Munc 18-like, Immunoglobulin-like and Type II Cytokeratin-like. More particularly, the invention relates to nucleic acids encoding novel polypeptides, as well as vectors, host cells, antibodies, and recombinant methods for producing these nucleic acids and polypeptides. Neuropilins play an active role in angiogenesis as they are receptors for VEGF and may regulate VEGF-induced angiogenesis. Neuropilins are also expressed by tumor cells and may play a role in tumor angiogenesis. Neuropilins also play a role in axon guidance as they bind to semaphorins and and in combination with plexins regulate the signal transduction events in neurons. Hence neuropilin-like molecules play an important role in embryonic development and the dedifferention events seen in cancer. These molecules probably play an important role in regulation of angiogenesis, cancer, development and neurological conditions. The MAM domain is characteristic ofthe extracellular region of membrane associated proteins such as meprin (a cell surface glycoprotein); A5 antigen (a developmentally-regulated cell surface protein); and receptor-like tyrosine protein phosphatase. These proteins although functionally diverse; resemble receptors comprising a signal peptide, an N-terminal extracellular domain, a single transmembrane domain and an intracellular domain. The MAM domain might play a role in cell adhesion.

Fibrillin is a very large molecule whose primary structure is now known from the cloning and sequencing of 10 kb of cDNA. Fibrillin is the major component of extracellular micro fibrils and is widely distributed in connective tissue throughout the body. Mutations in the fibrillin- 1 (FBN1) gene, on chromosome 15q21.1, have been found to cause Marfan syndrome, a dominantly inherited disorder characterised by clinically variable skeletal, ocular, and cardiovascular abnormalities. Fibrillin- 1 mutations have also been found in several other related connective tissue disorders, such as severe neonatal Marfan syndrome, dominant ectopia lentis, familial ascending aortic aneurysm, isolated skeletal features of Marfan syndrome, and Shprintzen-Goldberg syndrome (PMID: 9401003, PMLD: 8575254, PMID: 7584608).

Opioid binding cell adhesion molecules (OBCAMs) are members ofthe cell adhesion molecule family with homology to the immunoglobulin protein superfamily. OBCAMs are seen both in the developing nervous system as well as in the mature adult brain. They may play a role in neuronal outgrowth and development, probably by modulating cell-cell interactions. In addition, OBCAMs are known to affect the regulation and functioning of opioid receptors. Chronic morphine treatment downregulates the expression of at least one member of this family. Therefore, these proteins could mediate long-term effects on brain function by opioid usage and may be used as a therapeutic in that context.

The assimilation of dietary fats into the body requires that they be digested by Upases. One lipase, pancreatic triglyceride lipase, is essential for the efficient digestion of dietary fats. Pancreatic triglyceride lipase is the archetype ofthe lipase gene family that includes two homologues of pancreatic triglyceride lipase, pancreatic lipase-related proteins 1 and 2. The cDNA sequences encoding pancreatic triglyceride lipase and the related proteins have been described. Furthermore, the tertiary structure of human pancreatic triglyceride lipase has been determined alone and in a complex with colipase, a pancreatic protein required for lipase activity in the duodenum (Lowe, Molecular mechanisms of rat and human pancreatic triglyceride lipases. J Nutr 127(4):549-57, 1997).

The high-affinity receptor for immunoglobulin E, Fc epsilon RI, is found exclusively on mast cells and basophils. When multivalent allergens bind to the receptor-bound IgE, the consequent aggregation ofthe receptors leads to the release of mediators responsible for allergic symptoms. In rodents Fc epsilon RI is a tetrameric complex of non-covalently attached subunits: one IgE-binding alpha subunit, one beta subunit and a dimer of disulphide- linked gamma subunits (Blank et al., Complete structure and expression in transfected cells of high affinity IgE receptor. Nature 337(6203).T87-9, 1989). IgE receptors of mast cells, Fc epsilon RI, localize to coated pits and internalize after cross-linking. Studies have investigated whether any one ofthe receptor's four distinctive cytoplasmic domains regulates these phenomena. The conclusion of these studies was that no single cytoplasmic domain ofthe Fc epsilon RI uniquely controls its ligand-induced localization to coated pits and internalization (Mao et al., Effects of subunit mutation on the localization to coated pits and intemalization of cross-linked IgE-receptor complexes. J Immunol 151(5):2760-74, 1993).

Seel family proteins are regulators of diverse exocytic processes, from yeast to man. Three mammalian homologues, Muncl8-1, -2, and -3 have been described. The Muncl8-2 gene comprises 19 exons whose sizes range from 50 to 158 bp, with a total gene size of approximately 11 kb. A single transcript of 2.1 kb is expressed in multiple non-neuronal murine tissues. Muncl8-2 has a striking resemblance to Muncl8-1 in structure despite only 60% sequence identity, suggesting a recent gene duplication event (Agrawal et al., Gene structure and promoter function of murine Muncl8-2, a nonneuronal exocytic Seel homo log. Biochem Biophys Res Commun 276(3):817-22, 2000).

SUMMARY OF THE INVENTION

The invention is based in part upon the discovery of nucleic acid sequences encoding novel polypeptides. The novel nucleic acids and polypeptides are referred to herein as NOVX, or NOV1, NOV2, NOV3, NOV4, NOV5, NOV6, NOV7, NOV8, NOV9 and NOV10 nucleic acids and polypeptides. These nucleic acids and polypeptides, as well as derivatives, homologs, analogs and fragments thereof, will hereinafter be collectively designated as "NOVX" nucleic acid or polypeptide sequences.

In one aspect, the invention provides an isolated NOVX nucleic acid molecule encoding a NOVX polypeptide that includes a nucleic acid sequence that has identity to the nucleic acids disclosed in SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47. In some embodiments, the NOVX nucleic acid molecule will hybridize under stringent conditions to a nucleic acid sequence complementary to a nucleic acid molecule that includes a protein-coding sequence of a NOVX nucleic acid sequence. The invention also includes an isolated nucleic acid that encodes a NOVX polypeptide, or a fragment, homolog, analog or derivative thereof. For example, the nucleic acid can encode a polypeptide at least 80% identical to a polypeptide comprising the amino acid sequences of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48. The nucleic acid can be, for example, a genomic DNA fragment or a cDNA molecule that includes the nucleic acid sequence of any of SEQ LO NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47.

Also included in the invention is an oligonucleotide, e.g., an oligonucleotide which includes at least 6 contiguous nucleotides of a NOVX nucleic acid (e.g., SEQ LD NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47) or a complement of said oligonucleotide. Also included in the invention are substantially purified NOVX polypeptides (SEQ LD NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48). In certain embodiments, the NOVX polypeptides include an amino acid sequence that is substantially identical to the amino acid sequence of a human NOVX polypeptide. The invention also features antibodies that immunoselectively bind to NOVX polypeptides, or fragments, homologs, analogs or derivatives thereof.

In another aspect, the invention includes pharmaceutical compositions that include therapeutically- or prophylactically-effective amounts of a therapeutic and a pharmaceutically- acceptable carrier. The therapeutic can be, e.g., a NOVX nucleic acid, a NOVX polypeptide, or an antibody specific for a NOVX polypeptide. In a further aspect, the invention includes, in one or more containers, a therapeutically- or prophylactically-effective amount of this pharmaceutical composition.

In a further aspect, the invention includes a method of producing a polypeptide by culturing a cell that includes a NOVX nucleic acid, under conditions allowing for expression of the NOVX polypeptide encoded by the DNA. If desired, the NOVX polypeptide can then be recovered.

In another aspect, the invention includes a method of detecting the presence of a NOVX polypeptide in a sample. In the method, a sample is contacted with a compound that selectively binds to the polypeptide under conditions allowing for formation of a complex between the polypeptide and the compound. The complex is detected, if present, thereby identifying the NOVX polypeptide within the sample.

The invention also includes methods to identify specific cell or tissue types based on their expression of a NOVX.

Also included in the invention is a method of detecting the presence of a NOVX nucleic acid molecule in a sample by contacting the sample with a NOVX nucleic acid probe or primer, and detecting whether the nucleic acid probe or primer bound to a NOVX nucleic acid molecule in the sample.

In a further aspect, the invention provides a method for modulating the activity of a NOVX polypeptide by contacting a cell sample that includes the NOVX polypeptide with a compound that binds to the NOVX polypeptide in an amount sufficient to modulate the activity of said polypeptide. The compound can be, e.g., a small molecule, such as a nucleic acid, peptide, polypeptide, peptidomimetic, carbohydrate, lipid or other organic (carbon containing) or inorganic molecule, as further described herein. Also within the scope of the invention is the use of a therapeutic in the manufacture of a medicament for treating or preventing disorders or syndromes including, e.g., trauma, regeneration (in vitro and in vivo), viral/bacterial/parasitic infections, Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, Tuberous sclerosis, hypercalceimia, Parkinson's disease, Huntington's disease, Cerebral palsy, Epilepsy, Lesch-Nyhan syndrome, Multiple sclerosis, Ataxia-telangiectasia, Leukodystrophies, behavioral disorders, addiction, anxiety, pain, actinic keratosis, acne, hair growth diseases, allopecia, pigmentation disorders, endocrine disorders, connective tissue disorders, such as severe neonatal Marfan syndrome, dominant ectopia lentis, familial ascending aortic aneurysm, isolated skeletal features of Marfan syndrome, Shprintzen-Goldberg syndrome, genodermatoses, contractural arachnodactyly, inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer, leukemia or pancreatic cancer; blood disorders; asthma; psoriasis; vascular disorders, hypertension, skin disorders, renal disorders including Alport syndrome, immunological disorders, tissue injury, fibrosis disorders, bone diseases, Ehlers-Danlos syndrome type VI, VII, type IV, S-linked cutis laxa and Ehlers-Danlos syndrome type V, osteogenesis imperfecta, Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, immune disorders, hematopoietic disorders, muscle disorders, inflammation and wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, Treatment of Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, benign prostatic hypertrophy, arthrogryposis multiplex congenita, osteogenesis imperfecta, keratoconus, scoliosis, duodenal atresia, esophageal atresia, intestinal malrotation, Pancreatitis, Obesity Systemic lupus erythematosus,

Autoimmune disease, Emphysema, Scleroderma, allergy, ARDS, Neuroprotection, Fertility Myasthenia gravis, Diabetes, obesity, Growth and reproductive disorders Hemophilia, Hypercoagulation, Idiopathic thrombocytopenic purpura , Immunodeficiencies, Graft vesus host, Adrenoleukodystrophy , Congenital Adrenal Hyperplasia, Endometriosis, Xerostomia, Ulcers, Cirrhosis, Transplantation, Diverticular disease, Hirschsprung's disease, Appendicitis, Arthritis, Ankylosing spondylitis, Tendinitis, Renal artery stenosis, Interstitial nephritis, Glomerulonephritis, Polycystic kidney disease, erythematosus, Renal tubular acidosis, IgA nephropathy, anorexia, bulimia, psychotic disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease and/or other pathologies and disorders ofthe like.

The therapeutic can be, e.g., a NOVX nucleic acid, a NOVX polypeptide, or a NOVX- specific antibody, or biologically-active derivatives or fragments thereof. For example, the compositions ofthe present invention will have efficacy for treatment of patients suffering from the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. The polypeptides can be used as immunogens to produce antibodies specific for the invention, and as vaccines. They can also be used to screen for potential agonist and antagonist compounds. For example, a cDNA encoding NOVX may be useful in gene therapy, and NOVX may be useful when administered to a subject in need thereof. By way of non-limiting example, the compositions ofthe present invention will have efficacy for treatment of patients suffering from the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like.

The invention further includes a method for screening for a modulator of disorders or syndromes including, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. The method includes contacting a test compound with a NOVX polypeptide and determining if the test compound binds to said NOVX polypeptide. Binding ofthe test compound to the NOVX polypeptide indicates the test compound is a modulator of activity, or of latency or predisposition to the aforementioned disorders or syndromes. Also within the scope ofthe invention is a method for screening for a modulator of activity, or of latency or predisposition to disorders or syndromes including, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like by administering a test compound to a test animal at increased risk for the aforementioned disorders or syndromes. The test animal expresses a recombinant polypeptide encoded by a NOVX nucleic acid. Expression or activity of NOVX polypeptide is then measured in the test animal, as is expression or activity ofthe protein in a control animal which recombinantly- expresses NOVX polypeptide and is not at increased risk for the disorder or syndrome. Next, the expression of NOVX polypeptide in both the test animal and the control animal is compared. A change in the activity of NOVX polypeptide in the test animal relative to the control animal indicates the test compound is a modulator of latency ofthe disorder or syndrome.

In yet another aspect, the invention includes a method for determining the presence of or predisposition to a disease associated with altered levels of a NOVX polypeptide, a NOVX nucleic acid, or both, in a subject (e.g., a human subject). The method includes measuring the amount ofthe NOVX polypeptide in a test sample from the subject and comparing the amount ofthe polypeptide in the test sample to the amount ofthe NOVX polypeptide present in a control sample. An alteration in the level ofthe NOVX polypeptide in the test sample as compared to the control sample indicates the presence of or predisposition to a disease in the subject. Preferably, the predisposition includes, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. Also, the expression levels ofthe new polypeptides ofthe invention can be used in a method to screen for various cancers as well as to determine the stage of cancers.

In a further aspect, the invention includes a method of treating or preventing a pathological condition associated with a disorder in a mammal by administering to the subject a NOVX polypeptide, a NOVX nucleic acid, or a NOVX-specific antibody to a subject (e.g., a human subject), in an amount sufficient to alleviate or prevent the pathological condition. In preferred embodiments, the disorder, includes, e.g., the diseases and disorders disclosed above and/or other pathologies and disorders ofthe like. In yet another aspect, the invention can be used in a method to identity the cellular receptors and downstream effectors ofthe invention by any one of a number of techniques commonly employed in the art. These include but are not limited to the two-hybrid system, affinity purification, co-precipitation with antibodies or other specific-interacting molecules.

NOVX nucleic acids and polypeptides are further useful in the generation of antibodies that bind immuno-specifically to the novel NOVX substances for use in therapeutic or diagnostic methods. These NOVX antibodies may be generated according to methods known in the art, using prediction from hydrophobicity charts, as described in the "Anti-NOVX Antibodies" section below. The disclosed NOVX proteins have multiple hydrophilic regions, each of which can be used as an immunogen. These NOVX proteins can be used in assay systems for functional analysis of various human disorders, which will help in understanding of pathology ofthe disease and development of new drug targets for various disorders. The NOVX nucleic acids and proteins identified here may be useful in potential therapeutic applications implicated in (but not limited to) various pathologies and disorders as indicated below. The potential therapeutic applications for this invention include, but are not limited to: protein therapeutic, small molecule drug target, antibody target (therapeutic, diagnostic, drug targeting/cytotoxic antibody), diagnostic and/or prognostic marker, gene therapy (gene delivery/gene ablation), research tools, tissue regeneration in vivo and in vitro of all tissues and cell types composing (but not limited to) those defined here. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing ofthe present invention, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In the case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

Other features and advantages ofthe invention will be apparent from the following detailed description and claims.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel nucleotides and polypeptides encoded thereby. Included in the invention are the novel nucleic acid sequences and their encoded polypeptides. The sequences are collectively referred to herein as "NOVX nucleic acids" or "NOVX polynucleotides" and the corresponding encoded polypeptides are referred to as "NOVX polypeptides" or "NOVX proteins." Unless indicated otherwise, "NOVX" is meant to refer to any ofthe novel sequences disclosed herein. Table A provides a summary ofthe NOVX nucleic acids and their encoded polypeptides.

TABLE A. Sequences and Corresponding SEQ ID Numbers

NOVX nucleic acids and their encoded polypeptides are useful in a variety of applications and contexts. The various NOVX nucleic acids and polypeptides according to the invention are useful as novel members ofthe protein families according to the presence of domains and sequence relatedness to previously described proteins. Additionally, NOVX nucleic acids and polypeptides can also be used to identify proteins that are members ofthe family to which the NOVX polypeptides belong.

NOV1 is homologous to a Membrane Protein/ Neuropilin/Metalloproteinase-like family of proteins. Thus, the NOV1 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example; cancer, trauma, regeneration (in vitro and in vivo), viral/bacterial/parasitic infections, Von Hippel-Lindau (VHL) syndrome , Alzheimer's disease, stroke, Parkinson's disease, Huntington's disease, Cerebral palsy, Multiple sclerosis, hair growth diseases, endocrine disorders and/or other pathologies/disorders. NOV2 is homologous to a Fibrillin-like family of proteins. Thus NOV2 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example; connective tissue disorders, such as severe neonatal Marfan syndrome, Marfan syndrome, inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer, leukemia or pancreatic cancer; Neurologic diseases, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, inflammation, Parkinson's disease, osteoporosis, multiple sclerosis; angina pectoris, myocardial infarction, benign prostatic and/or other pathologies/disorders. NOV3 is homologous to a family of KIAAl 589-like proteins. Thus, the NOV3 nucleic acids and polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example: inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; skin disorders, Neurologic diseases, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, inflammation and wound repair; multiple sclerosis; angina pectoris, myocardial infarction, ulcers, benign prostatic hypertrophy and/or other pathologies/disorders.

NOV4 is homologous to the WD 40 motif-like family of proteins. Thus, NOV4 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated, for example; cancer, Diabetes, Von Hippel-Lindau (VHL) syndrome, Obesity Systemic lupus erythematosus , Autoimmune disease, Asthma, Von Hippel-Lindau (VHL) syndrome , Alzheimer's disease, Stroke,

Parkinson's disease, Huntington's disease, Cerebral palsy, Epilepsy, Lesch-Nyhan syndrome, Multiple sclerosis, Neuroprotection Fertility Myasthenia gravis, Neuroprotection Endocrine dysfunctions, obesity, Immunodeficiencies, Graft vesus host and/or other pathologies/disorders. NOV5 is homologous to the Opioid Bing Cell Adhesion Molecule-like protein family.

Thus NOV5 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example: Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalceimia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch- Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neurodegeneration and/or other pathologies/disorders.

NOV6 is homologous to the Triacylglycerol lipase-like family of proteins. Thus NOV6 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example: Diabetes, Von Hippel-Lindau (VHL) syndrome, Obesity, Von Hippel-Lindau (VHL) syndrome,

Transplantation, Inflammatory bowel disease, Arthritis, Alzheimer's disease, Stroke, Parkinson's disease, Huntington's disease, Endocrine dysfunctions, Diabetes, Growth and reproductive disorders, Multiple sclerosis, Autoimmune disease, Acne, Hair growth, allopecia, Hypercalceimia, Lesch-Nyhan syndrome and/or other pathologies/disorders. NOV7 is homologous to members ofthe IGE Receptor Beta Subunit-like family of proteins. Thus, the NOV7 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example; inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; asthma; psoriasis; skin disorders, renal disorders immunological disorders, bone diseases, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), anorexia, bulimia, asthma, osteoporosis, multiple sclerosis; benign prostatic hypertrophy, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease, Gilles de la Tourette syndrome and/or other pathologies/disorders.

NOV8 is homologous to the Munc 18-like family of proteins. Thus, NOV8 nucleic acids and polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example; inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; psoriasis; neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), Parkinson's disease, acute heart failure, hypotension, hypertension, osteoporosis, multiple sclerosis; angina pectoris, myocardial infarction, psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease, Gilles de la Tourette syndrome and/or other pathologies/disorders. NOV9 is homologous to members ofthe Immunoglobulin-like family of proteins. Thus, the NOV9 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example; inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; psoriasis; neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), Parkinson's disease, acute heart failure, hypotension, hypertension, osteoporosis, multiple sclerosis; angina pectoris, myocardial infarction, psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease, Gilles de la Tourette syndrome and/or other pathologies/disorders. NOV 10 is homologous to members ofthe Type II Cytokeratin-like family of proteins. Thus, the NOV 10 nucleic acids, polypeptides, antibodies and related compounds according to the invention will be useful in therapeutic and diagnostic applications implicated in, for example; inflammatory and infectious diseases such as AIDS; Neurologic diseases, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), anorexia, bulimia, asthma, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, Crohn's disease; multiple sclerosis; and Treatment of Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, asthma, allergies, benign prostatic hypertrophy, and psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease and/or other pathologies/disorders. The NOVX nucleic acids and polypeptides can also be used to screen for molecules, which inhibit or enhance NOVX activity or function. Specifically, the nucleic acids and polypeptides according to the invention may be used as targets for the identification of small molecules that modulate or inhibit, e.g., neurogenesis, cell differentiation, cell proliferation, hematopoiesis, wound healing and angiogenesis. Additional utilities for the NOVX nucleic acids and polypeptides according to the invention are disclosed herein.

NOV1

NOV1 includes two novel Membrane protein neuropilin/metalloproteinase-like proteins disclosed below. The disclosed proteins have been named NOV1 a and NOVlb. NOVla

A disclosed NOVla nucleic acid of 1668 nucleotides (also referred to as SC40376139) encoding a novel Membrane protein/neuropilin/metalloproteinase-like protein is shown in Table 1 A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 7-9 and ending with a TGA codon at nucleotides 1453-1455. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 1 A, and the start and stop codons are in bold letters. Table 1A. NOVla Nucleotide Sequence (SEQ ID NO:l).

GGATCΆATGCAAATGGAGΆGTTATGATGGAACACTGAGGATTGTGAATGTATCTAGGGAAATGTCAGGAATGTACAGATGT CAGACCAGCCAATACAATGGATTTAACGTGAAACCAAGGGAAGCCTTGGTGCAGCTCATCGTTCAGTATCCCCCTGCAGTG GAACCAGCATTCTTGGAAATCCGGCAAGGACAGGATCGAAGTGTCACTATGAGTTGCAGAGTACTGAGAGCCTATCCAATA CGGGTGCTGACCTATGAGTGGCGCTTGGGCAATAAATTATTACGGACGGGTCAATTTGACTCTCAGGAATACACAGAGTAC GCTGTGAAGAGTCTTTCCAATGAAAACTATGGGGTTTATAACTGTAGCATCATAAATGAAGCTGGAGCTGGGAGATGCAGC TTTCTTGTTACAGGTGGAAAGGCCTATGCTCCAGAATTCTATTATGATACCTACAATCCAGTATGGCAGAACAGACACCGT GTTTATTCTTACAGTCTACAGTGGACACAGATGAATCCTGATGCAGTGGATCGGATTGTTGCATACCGGTTGGGCATCAGG CAGGCTGGACAGCAGCGCTGGTGGGAGCAGGAGATTAAAATAAATGGGAATATTCAAAAGGGAGAATTAATTACATATAAC TTGACAGAGCTAATTAAACCAGAAGCTTATGAAGTCCGACTGACTCCTCTCACCAAATTTGGTGAAGGAGATTCAACAATT CGTGTGATCAAATATAGTCCTGTAAATCCTCATTTGAGTGAATTTCATTGTGGATTTGAAGATGGTAATATTTGTTTGTTC ACTCAAGATGATACAGATAATTTTGACTGGACAAAGCAAAGTACAGCAACAAGAAATACAAAATATACTCCTAATACAGGA CCTAATGCTGACCGTAGTGGCTCCAAAGAAGGTTTTTATATGTACATTGAGACATCACGACCCAGATTGGAAGGCGAAAAG GCTCGACTTCTCAGCCCTGTTTTCAGCATAGCTCCAGCCCTGTTTTCAGCAAGACTTCTCAGCCCTGTTTTCAGCATAGCT CCCAAAAACCCTTATGGACCCACAAACACTGCATATTGTTTCAGCTTCTTTTATCACATGTATGGACAACATATAGGTGTC TTAAATGTTTATCTACGTTTGAAAGGGCAAACAACAATAGAGAATCCACTGTGGTCTTCAAGTGGGAATAAAGGACAAAGA TGGAATGAGGCTCATGTTAATATATACCCAATTACTTCATTTCAGCTCATTTTTGAAGGTATCCGAGGTCCTGGAATAGAA GGTGACATTGCTATTGATGATGTATCAATTGCAGAAGGAGAATGTGCAAAACAAGACCTAGCAACTAAGAATTCCGTTGAT GGTGCTGTTGGGATTTTGGTTCATATATGGCTTTTTCCCATTATCGTCCTCATCTCTATCTTAAGTCCTCGAAGGTGACCT TATCCTGGCAGAGGCTATAAAAGATTCACCAGGCACTGGCATGAAGAAAGAGTCTTTGTAAATGGACATTGAACAAACAAA CTACCAAAGATTCCTCCACTGACTΆCTGACTCAAAAATAAAATAATAAAAACAAATTTTTTTAAGCACTGGGGATAAAAΆG ACATCATGGAAGTATAACTTATTCAGACTAAΆCATAAAGATAΆTCTGA

The NOVla nucleic acid sequence maps to chromosome 14 and has 1030 of 1128 bases (91%) identical to a Macacafascicularis brain mRNA (gb:GENBANK- ID:AB047834|acc:AB047834.1) (E = 3.2e^"201). Similiarity information was assessed using public nucleotide databases including all GenBank databases and the GeneSeq patent database. Chromosome information was assigned using OMIM and the electronic northern tool from Curatools to derive the the chromosomal mapping ofthe SeqCalling assemblies, Genomic clones, and/or EST sequences that were included in the invention.

In all BLAST alignments herein, the "E-value" or "Expect" value is a numeric indication ofthe probability that the aligned sequences could have achieved their similarity to the BLAST query sequence by chance alone, within the database that was searched. For example, the probability that the subject ("Sbjct") retrieved from the NOVla BLAST analysis, e.g., Macacafascicularis brain mRNA, matched the Query NOVla sequence purely by chance is 3.2e^" . The Expect value (E) is a parameter that describes the number of hits one can "expect" to see just by chance when searching a database of a particular size. It decreases exponentially with the Score (S) that is assigned to a match between two sequences. Essentially, the E value describes the random background noise that exists for matches between sequences.

The Expect value is used as a convenient way to create a significance threshold for reporting results. The default value used for blasting is typically set to 0.0001. In BLAST 2.0, the Expect value is also used instead ofthe P value (probability) to report the significance of matches. For example, an E value of one assigned to a hit can be interpreted as meaning that in a database ofthe current size one might expect to see one match with a similar score simply by chance. An E value of zero means that one would not expect to see any matches with a similar score simply by chance. See, e.g., http://www.ncbi.nlm.nih.gov/Education/BLASTinfo/. Occasionally, a string of X's or N's will result from a BLAST search. This is a result of automatic filtering ofthe query for low- complexity sequence that is performed to prevent artifactual hits. The filter substitutes any low-complexity sequence that it finds with the letter "N" in nucleotide sequence (e.g., "NN^N NNN") or the letter "X" in protein sequences (e.g., "XXX"). Low-complexity regions can result in high scores that reflect compositional bias rather than significant position- by-position alignment. Wootton and Federhen, Methods Enzymol 266:554-571, 1996.

The disclosed NOVla polypeptide (SEQ ID NO:2) encoded by SEQ ID NO:l has 482 amino acid residues and is presented in Table IB using the one-letter amino acid code. Signal P, Psort and/or Hydropathy results predict that NOVla does not contain a signal peptide and is likely to be localized to the plasma membrane with a certainty of 0.7000.

Table IB. Encoded NOVla protein sequence (SEQ ID NO:2). QMESYDGTLRIVNVSRE SGMYRCQTSQYNGFNVKPREALVQLIVQYPPAVEPAFLEIRQGQDRSVTMSCRVLRAYPIRV LTYE RLGNKLLRTGQFDSQEYTEYAVKSLSNENYGVYNCSIINEAGAGRCSFLVTGGKAYAPEFYYDTYNPVWQNRHRVY SYSLQWTQMNPDAVDRIVAYR GIRQAGQQRWWEQEIKINGNIQKGE ITYNLTE IKPEAYEVRLTPLTKFGEGDSTIRV IKYSPVNPHLΞEFHCGFEDGNICLFTQDDTDNFDWTKQSTATRNTKYTPNTGPNADRSGSKEGFYMYIETSRPRLEGEKAR SPVFSIAPALFSARLLSPVFSIAPKNPYGPTNTAYCFSFFYHMYGQHIGVLNVY RLKGQTTIENPLWSSΞGNKGQR N EAHVNIYPITSFQLIFEGIRGPGIEGDIAIDDVSIAEGECAKQDLATKNSVDGAVGILVHIWLFPIIVLIΞI SPRR

The NOVla amino acid sequence has 445 of 464 amino acid residues (95%) identical to, and 445 of 464 amino acid residues (95%) similar to, a Macacafascicularis 448 amino acid residue hypothetical 51.2 kda protein (ptnr:TREMBLNEW-ACC:BAB 12260) (E = l.Oe^" ). The NOVla amino acid sequence also has 71 of 197 amino acid residues (36%) identical to, and 108 of 197 amino acid residues (55%) similar to, aXenopus lαevis 928 amino acid residue neuroρilin-1 precursor protein (ptnr:SWISSNEW-ACC:P28824) (E = l.Oe^"17). NOVla is predicted to be expressed in brain tissues because ofthe expression pattern of a closely related Macaca fascicularis brain cDNA, clone:QccE-16296 homolog (GENBANK-ID: gb:GENBANK-ID: AB047834|acc: AB047834.1).

NOVlb

A disclosed NOVlb nucleic acid of 1608 nucleotides (also referred to as CG55014-02) encoding a novel Membrane protein/neuropilin/metalloproteinase-like protein is shown in Table lC. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 6-8 and ending with a TGA codon at nucleotides 1404-1406. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 1C, and the start and stop codons are in bold letters. Table lC. NOVlb Nucleotide Sequence (SEQ ID NO:3).

GATCAATGCAAATGGAGAGTTATGΆTGGAACΆCTGAGGATTGTGAATGTATCTAGGGAAATGTCAGGAATGTΆCAGATGTC AGACCAGCCAATACAATGGATTTAACGTGAAACCAAGGGAAGCCTTGGTGCAGCTCATCGTTCAGTATCCCCCTGCAGTGG AACCAGCATTCTTGGAAATCCGGCAAGGACAGGATCGAAGTGTCACTATGAGTTGCAGAGTACTGAGAGCCTATCCAATAC GGGTGCTG_ACCTATGAGTGGCGCTTGGGCAATAAATTATTACGGACGGGTCAATTTGACTCTCAGGAATACACAGAGTACG CTGTGAAGAGTCTTTCCAATGAAAACTATGGGGTTTATAACTGTAGCATCATAAATGAAGCTGGAGCTGGGAGATGCAGCT TTCTTGTTACAGGAAAGGCCTATGCTCCAGAATTCTATTATGATACCTACAATCCAGTATGGCAGAACAGACACCGTGTTT ATTCTTACAGTCTACAGTGGACACAGATGAATCCTGATGCAGTGGATCGGATTGTTGCATACCGGTTGGGCATCAGGCAGG CTGGACAGCAGCGCTGGTGGGAGCAGGAGATTAAAATAAATGGGAATATTCAAAAGGGAGAATTAATTACATATAACTTGA CAGAGCTAATTAAACCAGAAGCTTATGAAGTCCGACTGACTCCTCTCACCAAATTTGGTGAAGGAGATTCAACAATTCGTG

TGATCAAATATAGTGCTCCTGTAAATCCTCATTTGAGAGAATTTCATTGTGGATTTGAAGATGGTAATATTTGTTTGTTCA CTCAAGATGATACAGATAATTTTGACTGGACAAAGCAAAGTACAGCAACAAGAAATACAAAATATACTCCTAATACAGGAC CTAATGCTGACCGTAGTGGCTCCAAAGAAGGTTTTTATATGTACATTGAGACATCACGACCCAGATTGGAAGGCGAAAAGG CTCGACTTCTCAGCCCTGTTTTCAGCATAGCTCCCAAAAACCCTTATGGACCCACAAACACTGCATATTGTTTCAGCTTCT TTTATCACATGTATGGACAACATATAGGTGTCTTAAATGTTTATCTACGTTTGAAAGGGCAAACAACAATAGAGAATCCAC TGTGGTCTTCAAGTGGGAATAAAGGACAAAGATGGAATGAGGCTCATGTTAATATATACCCAATTACTTCATTTCAGCTCA TTTTTGAAGGTATCCGAGGTCCTGGAATAGAAGGTGACATTGCTATTGATGATGTATCAATTGCAGAAGGAGAATGTGCAA AACAAGACCTAGCAACTAAGAATTCCGTTGATGGTGCTGTTGGGATTTTGGTTCATATATGGCTTTTTCCCATTATCGTCC TCACCTCTATCTTAAGTCCTCGAAGGTGACCTTATCCTGGCAGAGGCTATAAAAGATTCACCAGGCACTGGCATGAAGAAA GAGTCTTTGTAAATGGACATTGAACAAACAAACTACCAAAGATTCCTCCACTGACTACTGACTCAAAAATAAAATAATAAA AACAAATTTTTTTAAGCACTGGGGATAAAAAGACATCATGGAAGTATAACTTATTCAGACTAAACATAA

The NOVlb nucleic acid sequence has 1538 of 1557 bases (98%) identical to a Macacafascicularis brain mRNA (gb:GENBANK-ID:AB047834|acc:AB047834.1) (E = 0.0).

The disclosed NOVlb polypeptide (SEQ ID NO:4) encoded by SEQ ID NO:3 has 466 amino acid residues and is presented in Table ID using the one-letter amino acid code. Signal P, Psort and/or Hydropathy results predict that NOVlb does not contain a signal peptide and is likely to be localized to the plasma membrane with a certainty of 0.7000.

Table ID. Encoded NOVlb protein sequence (SEQ ID NO:4).

MQMESYDGTLRIVNVSREMSGMYRCQTSQYNGFNVKPREALVQLIVQYPPAVEPAFLEIRQGQDRSVTMSCRV RAYPIRV LTYEWRLGNKLLRTGQFDSQEYTEYAVKSLSNENYGVYNCSIINEAGAGRCSFLVTGKAYAPEFYYDTYNPVWQNRHRVYS YSLQWTQMNPDAVDRIVAYRLGIRQAGQQRWWEQEIKINGNIQKGELITYNLTELIKPEAYEVRLTPLTKFGEGDSTIRVI KYSAPVNPHLREFHCGFEDGNICLFTQDDTDNFDWTKQSTATRNTKYTPNTGPNADRSGSKEGFYMYIETSRPRLEGEKAR LLSPVFSIAPKNPYGPTNTAYCFSFFYHMYGQHIGVLNVY RLKGQTTIENP SSSGNKGQRWNEAHVNIYPITSFQLIF EGIRGPGIEGDIAIDDVΞIAEGECAKQDLATKNSVDGAVGILVHIWLFPIIVLTSI SPRR

The NOVlb amino acid sequence has 446 of 448 amino acid residues (99%) identical to, and 446 of 448 amino acid residues (99%) similar to, a Macacafascicularis 448 amino acid residue hypothetical 51.2 kda protein (ptnr:TREMBLNEW-ACC:BAB12260) (E = 3.1e^"

248

NOVlb is expressed in at least the following tissues: brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Literature sources, and/or RACE sources. In addition, NOVlb is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Macaca fascicularis brain cDNA, clone:QccE- 16296 homolog (GENBANK-ID: gb:GENBANK- ιD:AB047834|acc:AB047834.1):Brain. Possible small nucleotide polymorphisms (SNPs) found for NOVlb are listed in Tables IE and IF. Depth represents the number of clones covering the region ofthe SNP. The putative allele frequence (PAF) is the fraction of these clones containing the SNP. A dash, when shown, means that a base is not present. The sign ">" means "is changed to."

NOVla and NOVlb are very closely homologous as is shown in the amino acid alignment in Table 1 G.

Table 1G Amino Acid Alignment of NOVla and NOVlb

NOVla NOVlb

110 120 130 1 0 150 I I ..

NOVla )EYTEYAVKS SNENYGVYNCSII EAGAGRC !R3|3aEπ NOVlb )EYTEYAVKSLSNENYGVYNCSIINEAGAGRCSF

160 170 180 190 200 . . I . .

NOVla NOVlb 'NPVWQNRHRVYSYS OWTQMNPDAVDRIVAYRLGI RQAGOQRWWEOE

210 220 230 240 250

. . I . . • • I

NOVla NOVlb ππrajMaMrøMiiBB

260 270 280 290 300

NOVla EFHCGFEDGNIC FTQDDTDNFDWTKQSTATRNTKYTPNTGPNA NOVlb EFHCGFEDGNICLFTQDDTDNFDWTKQSTATRNTKYTPNTGPNA

310 320 330 340 350

NOVla RSGSKEGFYMYIETSRPRLEGEKARLLS IPVFS I APALFSARLLSR NOVlb RSGSKEGFYMYIETSRPRLEGEKAR S NOVla NOVlb

410 420 430 440 450

NOVla ix nax NOVlb BiBwaw HwaBuHwtff IE tww

NOVla NOVlb

Homologies to any ofthe above NOV1 proteins will be shared by the other NOV1 proteins insofar as they are homologous to each other as shown above. Any reference to NOV1 is assumed to refer to both ofthe NOV1 proteins in general, unless otherwise noted.

NOVla also has homology to the amino acid sequences shown in the BLASTP data listed in Table IH.

The homology between these and other sequences is shown graphically in the ClustalW analysis shown in Table II. In the ClustalW alignment ofthe NOVla protein, as well as all other ClustalW analyses herein, the black outlined amino acid residues indicate regions of conserved sequence (i.e., regions that may be required to preserve structural or functional properties), whereas non-highlighted amino acid residues are less conserved and can potentially be altered to a much broader extent without altering protein structure or function.

Table II. ClustalW Analysis of NOVla

1) Novel NOVla (SEQ ID NO:2)

2) ^gil9967117ldbilB AB12260.il (AB047834) hypothetical protein [Macaca fascicularis] (SEQ ID NO:49)

3) ^gil7529597lemblCAB86653.11 (AL049553) dJ402N21.2 (novel protein with MAM domain) [Homo sapiens] (SEQ ID NO:50)

4) gi|12836077|dbι|BAB23491.1| (AK004706) putative [Mus musculus] (SEQ ID NO:51)

5) gi|4929103|gb[AAD33860.1IAF140020 1 (AF 140020) metalloproteinase 2 [Hydra vulgaris] (SEQ ID NO:52)

6) gi|15929977|gblAAH15417.1|AAH15417 (BC015417) Unknown (protein for MGC:21981) [Homo sapiens]

(SEQ ID NO:53)

10 20 30 40 50 ....|.... |.... |.... I.... I.... I...._ .... I.... I.... I

NOVla MQ ESYDGTLRIVNVSREMSGMYRCfflTSQ NfflFN KPRHALVQlinQYPP gl I 9967117 | -MSG YRCiTSO NBFNJJKPRgALVQllHQYPP gi 17529597 I gil 12836077 I gi I 4929103 I TKLVLIL ΞVALCHSFPEEENNDPBGIL1G@DIILTP@QKSI1EBGGDI gi 1159299771

70 80 90 100

110 120 130 140 150 I I I I I I I .... I I I

NOVla QEYTEYAVKΞffl|N|NYGVYNCSHlNEAg3fflRCB fflvTGGKAYAPEFYYDT gi| 9967117 | QEYTEYPVKsHsNENYGVYNCsHlNEAH BRcH|fflvTG-KAYAPEFYYDT gi I 7529597 I gi|12836077| gι|4929103| AFRE EERSCJJ|E RRTDEKDY(3EFFQ@lScW@f|jGRVGGLQNIS DDGC gi| 15929977 I

160 170 180 190 200

The presence of identifiable domains in NOVla, as well as all other NOVX proteins, was determined by searches using software algorithms such as PROSITE, DOMAIN, Blocks, Pfam, ProDomain, and Prints, and then determining the Interpro number by crossing the domain match (or numbers) using the Interpro website (http:www.ebi.ac.uk/ interpro). DOMAIN results for NOVla, as disclosed in Tables 1 J and IK, were collected from the Conserved Domain Database (CDD) with Reverse Position Specific BLAST analyses. This BLAST analysis software samples domains found in the Smart and Pfam collections. For Tables 1 J, IK and all successive DOMAIN sequence alignments, fully conserved single residues are indicated by black shading or by the sign (|) and "strong" semi-conserved residues are indicated by grey shading or by the sign (+). The "strong" group of conserved amino acid residues may be any one ofthe following groups of amino acids: STA, NEQK, NHQK, NDEQ, QHRK, MILV, MILF, HY, FYW. Tables 1J and IK lists the domain description from DOMAIN analysis results against

NOVla. This indicates that the NOVla sequence has properties similar to those of other proteins known to contain these domains.

Table 1J. Domain Analysis of NOVla gnl I Smart I smart00137, MAM, Domain in meprin, A5, receptor protein tyrosine phosphatase u (and others); Likely to have an adhesive function. Mutations in the meprin MAM domain affect noncovalent associations within meprin oligomers. In receptor tyrosine phosphatase mu-like molecules the MAM domain is important for homophilic cell-cell interactions. (SEQ ID NO: 54)

Length = 163 residues, 99.4% aligned

Score = 154 bits (390) , Expect = 9e-39 NOVla 254 SEFHCGFEDGNICLFTQDDTDNFDWTKQSTATRNTKYTPNTGPNADRSGSKEGFYMYIET 313 1+ +1 I l + l I I + I I 1+ I + l + l I I I I I I + I 11+ I I 00137 2 SDGNCDFEEGNTCGWHQDSNDDGP ERVSSAT RNDGPNRDHTT-GNGHYMFFET 54

NOVla 314 SRPRLEGEKARLLSPVFSIAPALFSARLLSPVFSIAPKNPYGPTNTAYCFSFFYHMYGQH 373

I 1+ I I I I I I + +1 +1 + 1 + 1 I I

00137 55 S-SGKPGQTARLLSPPLY ENRSTHCLTFWYYMYGSG 89

NOVla 374 IGVLNVYLRLKGQTTIENPL SSSGNKGQRWNEAHVNIYPITS-FQLIFEGIRGPGIEGD 432

+ 1 I I I 1 + 1+ + III II +1 +1 +1 1 + I I++I I I I I I I 00137 90 VGTLNVYVRVNN-GPQDTLLWSRSGTQGGQWLQAEVALSTSPQPFQWFEGTRGGGPSGY 148

NOVla 433 IAIDDVSIAEGECAK 447

I I + II + ++ I I I 00137 149 IALDDILLSNGPCGK 163

Table IK. Domain Analysis of NOVla qnl I Smart I smart00230, CysPc, Calpain-like thiol protease family. ; Calpain-like thiol protease family (peptidase family C2) . Calcium activated neutral protease (large subunit). (SEQ ID NO: 55) Length = 323 residues, 99.1% aligned Score = 342 bits (877), Expect = 4e-95

NOVla 258 CGFEDGNICLFTQDDTDNFDWTKQSTATRNTKYTPNTGPNADRSGSKEGFYMYIETSRPR 317

I I I I 1+ I ++ 1 I 1+ I I 1+ ++I I + III I + I I I I++I I 00230 1 CDFEDGSHCG SQDSGDDLD TRVNSATGGS TGPRGDHTT-GNGHYMYVDTS-SG 53

NOVla 318 LEGEKARLLSPVFSIAPALFSARLLSPVFSIAPKNPYGPTNTAYCFSFFYHMYGQHIGVL 377

I 1+ I I I I I I l + l +1 + 1 llll +1 1

00230 54 QEGQTARLLSPPLP PKRSPCCLTF YHMYGSGVGTL 89

NOVla 378 NVYLRLKGQTTIENPLWSSSGNKGQRWNEAHVNIYPITS-FQLIFEGIRGPGIEGDIAID 436

I I l + l + + I I I I I++I I 1 1 + I I I++I I I I I I I I l + l 00230 90 NVYVR-ENGGPSDRLLWSRSGHQGGSWLLAEVTLPTΞTKPFQWFEGTRGGGSRGGIALD 148

NOVla 437 DVSIAEGECAK 447

I+I++I I I + 00230 149 DISLSEGPCNQ 159

Recent immunohistochemistry studies have attemted to examine the expression of VEGF isoforms and their receptors in fibrovascular tissues obtained from proliferative diabetic retinopathy cases. RT-PCR analysis demonstrated the expression of VEGF receptors VEGF- Rl, VEGF-R2, and neuropilin-1 in 12, 14, and 14 of 22 proliferative diabetic retinopathy cases, respectively. Notably, VEGF-R2 and neuropilin-1 were simultaneously expressed in the identical 14 tissues. The isoform VEGF121 was constitutively expressed in all the tissues examined, whereas the expression of VEGF165 was confined to the 7 tissues that also expressed VEGF-R2 and neuropilin-1. The vascular density of fibrovascular tissues evaluated by immunohistochemistry for CD34 was significantly higher in the cases with the expression of VEGF-R2 and neuropilin-1 than in those without their expression (P < 0.01), whereas VEGF-R1 expression had no such relationship with the vascular density. The fibrovascular tissues that expressed VEGF 165 together with VEGF-R2 and neuropilin-1 were found in significantly younger patients (P < 0.01). In situ hybridization and immunohistochemical studies demonstrated that glial cells in the fibrovascular tissues express and produce VEGF. Coexpression of VEGF-R2 and neuropilin-1 is suggested to facilitate fibrovascular proliferation in diabetic retinopathy (Ishida et al., Coexpression of VEGF receptors VEGF-R2 and neuropilin-1 in proliferative diabetic retinopathy. Invest Ophthalmol Vis Sci 41(7): 1649- 56, 2000.)

In the developing nervous system axons navigate with great precision over large distances to reach their target areas. Chemorepulsive signals such as the semaphorins play an essential role in this process. The effects of one of these repulsive cues, semaphorin 3 A (Sema3A), are mediated by the membrane protein neuropilin-1 (Npn-1). Recent work has shown that neuropilin-1 is essential but not sufficient to form functional Sema3A receptors and indicates that additional components are required to transduce signals from the cell surface to the cytoskeleton. It was shown that members of the plexin family interact with the neuropilins and act as co-receptors for Sema3 A. Neuropilin/plexin interaction restricts the binding specificity of neuropilin-1 and allows the receptor complex to discriminate between two different semaphorins. Deletion ofthe highly conserved cytoplasmic domain of Plexin-Al or -A2 creates a dominant negative Sema3A receptor that renders sensory axons resistant to the repulsive effects of Sema3A when expressed in sensory ganglia. These data suggest that functional semaphorin receptors contain plexins as signal-transducing and neuropilins as ligand-binding subunits (Rohm et al., Plexin/neuropilin complexes mediate repulsion by the axonal guidance signal semaphorin 3A. Mech Dev 93(l-2):95-104, 2000).

Mesangial cell proliferation and growth factor over-expression are characteristic features of several glomerular diseases. Vascular endothelial growth factor (VEGF), a potent mitogen, is expressed in podocytes in the glomerulus, and VEGF receptors (flt-1, KDR, and neuropilin-1) are present on endothelial cells and other cell types. In conclusion, flt-1, KDR, and neuropilin-1 are present on cultured HMC, and VEGF(165) induces HMC proliferation. In addition, the flt-1 and KDR receptors are expressed in the mesangium in mesangioproliferative disease (Thomas et al., Vascular endothelial growth factor receptors in human mesangium in vitro and in glomerular disease. J Am Soc Nephrol 2000 l l(7):1236-43, 2000).

Vascular endothelial growth factor (VEGF), a major regulator of angiogenesis, binds to two receptor tyrosine kinases, KDR/Flk-1 and Flt-1. A third VEGF receptor, one that binds

VEGF 165 but not VEGF121, was cloned and purified from tumor cells. This isoform-specific VEGF receptor (VEGF165R) is identical to human neuropilin-1, a receptor for the collapsin/semaphorin family that mediates neuronal cell guidance. When coexpressed in cells with KDR, neuropilin-1 enhances the binding of VEGF165 to KDR and VEGF165-mediated chemotaxis. Conversely, inhibition of VEGF165 binding to neuropilin-1 inhibits its binding to KDR and its mitogenic activity for endothelial cells. It is proposed that neuropilin-1 is a novel VEGF receptor that modulates VEGF binding to KDR and subsequent bioactivity and therefore may regulate VEGF-induced angiogenesis (Soker et al., Neuropilin-1 is expressed by endothelial and tumor cells as an isoform-specific receptor for vascular endothelial growth factor. Cell 92(6):735-45, 1998).

The above defined information for NOV1 suggests that this Membrane protein/neuropilin/metalloproteinase-like protein may function as a member ofthe Membrane protein/neuropilin/metalloproteinase family. Therefore, the NOV1 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV1 compositions ofthe present invention will have efficacy for treatment of patients suffering from cancer, trauma, regeneration (in vitro and in vivo), viral/bacterial/parasitic infections, Von Hippel-Lindau (VHL) syndrome , Alzheimer's disease, stroke, Tuberous sclerosis, hypercalceimia, Parkinson's disease, Huntington's disease, Cerebral palsy, Epilepsy, Lesch- Nyhan syndrome, Multiple sclerosis, Ataxia-telangiectasia, Leukodystrophies, behavioral disorders, addiction, anxiety, pain, psoriasis, actinic keratosis , acne, hair growth diseases, allopecia, pigmentation disorders and endocrine disorders. The NOV1 nucleic acid encoding Membrane protein/neuropilin/metalloproteinase-like protein, and the Membrane protein/neuropilin/metalloproteinase-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV2

NOV2 includes six novel Fibrillin-like proteins disclosed below. The disclosed proteins have been named NOV2a, NOV2b, NOV2c, NOV2d, NOV2e and NOV2f.

NOV2a

A disclosed NOV2a nucleic acid of 9993 nucleotides (also referred to as GMAC022146_A) encoding a novel Fibrillin-like protein is shown in Table 2 A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 166-168 and ending with a TGA codon at nucleotides 9151-9153. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table 2A. The start and stop codons are in bold letters. Table 2A. NOV2a nucleotide sequence (SEQ ID NO:5).

CCTCCCATTCCTCTCCTCTCTCTCCCATCTGGACAGCCCCCAGCCTTCTGACACCCTGTTTCCCCCCCGGGACCCTGCTGC CCTTCCCCACACCACCTAACATGCATTTCTGACCTTGCTCCTCCCACTCAAAGCTTTTCAAGGGCTCTTCATTGTCCTTGA CGAATGAGCAAAAGCTGCATGACACTGCTATGCCCACTCCTGCAACGCCTGGCCCCACCTGGCCCCACCTGGCCCTCTCTC CCCTCCCTTCCACTTCCCACTCCCGTCTTGCCTTGGCTGTGCACCCTCAGCCCCAAGACCCTTCTCTAACATCACACCTCC TCCAGGAAGCCTCCTCTGACTGTTCACCGCTTCCCGGTTGGGGCTGGATGCCTCCTTGGGCTCCCAGAGGCCCCGGGGCTG CCCCATTTGGCAACCACCCTGCCCTCCTCGTTGTGTTGGGTCCCCCATGGCTAGGGTGGGGGGCTCATCTAGGGCTGACTC TGCTTCCTCTCCGCAGCCTCCAGGGGACACGCCATGACTCTGGAGGGTCTGTATTTGGCAAGGGGCCCCCTGGCCCGGCTC CTGCTGGCCTGGTCGGCCCTGTTGTGCATGGCAGGTGGCCAAGGCCGCTGGGACGGGGCCTTGGAGGCTGCAGGTCCTGGA CGTGTGCGGAGGCGGGGCAGCCCAGGCATCTTGCAGGGGCCGAATGTGTGCGGCTCCCGGTTCCATGCCTACTGCTGTCCA GGCTGGAGGACATTCCCTGGCAGGAGCCAGTGTGTCGTACCCATCTGTAGGCGCGCCTGCGGTGAAGGCTTCTGCTCCCAG CCCAACCTGTGCACCTGTGCGGATGGGACGCTGGCTCCCAGCTGCGGGGTGAGCCGAGGGTCAGGGTGCAGTGTGAGCTGT ATGAATGGGGGCACCTGCCGGGGGGCGTCCTGTCTGTGTCAGAAGGGCTACACAGGCACCGTGTGTGGGCAGCCCATCTGT GACCGCGGCTGCCACAATGGGGGTCGCTGCATTGGGCCCAACCGCTGCGCCTGTGTGTATGGCTTCATGGGACCTCAATGT GAGAGAGATTACCGGACGGGACCCTGCTTTGGCCAAGTAGGCCCCGAGGGGTGCCAGCATCAGCTGACGGGCCTCGTGTGC ACCAAGGCACTTTGCTGTGCCACTGTGGGCCGTGCCTGGGGCCTTCCATGTGAACTTTGCCCTGCACAGCCACACCCCTGC CGCCGCGGCTTCATCCCCAATATCCACACGGGGGCCTGCCAAGATGTGGATGAGTGCCAGGCTGTGCCAGGCCTGTGCCAG GGAGGCAGCTGCGTCAACATGGTGGGCTCCTTCCATTGCCGCTGTCCAGTTGGACACCGGCTCAGTGACAGCAGCGCCGCA TGTGAAGACTACGATGAATGCAGCACCATTCCTGGAATCTGTGAAGGGGGTGAATGTACAAACACAGTCAGCAGTTACTTT TGCAAATGTCCTCCTGGTTTTTACACCTCTCCAGATGGTACTCTTCATGGACAGTCGCGGGCCGGCGCCTGCTTCTCAGTG CTTTTCGGGGGCCGCTGTGCTGGAGACCTCGCCGGCCACTACACTCGCAGGCAGTGCTGCTGTGACAGGGGCAGGTGCTGG GCAGCTGGCCCGGTCCCTGAGCTGTGTCCTCCTCGGGGCTCCAATGAATTCCAGCAACTGTGCGCCCAGCGGCTGCCGCTG CTACCCGGCCACCCTGGCCTCTTCCCTGGCCTCCTGGGCTTCGGATCCAATGGCATGGGTCCCCCTCTTGGGCCAGCGCGA CTCAACCCCCATGGCTCTGATGCGCGTGGGATCCCCAGCCTGGGCCCTGGCAACTCTAATATTGGCACTGCTACCCTGAAC CAGACCATTGACATCTGCCGACACTTCACCAACCTGTGTCTGAATGGCCGCTGCCTGCCCACGCCTTCCAGCTACCGCTGC GAGTGTAACGTGGGCTACACCCAGGACGTGCGCGGCGAGTGCATTGATGTAGACGAATGCACCAGCAGCCCCTGCCACCAC GGTGACTGCGTCAACATCCCCGGCACCTACCACTGCCGGTGCTACCCGGGCTTCCAGGCCACGCCCACCAGGCAGGCATGC GTGGATGTGGACGAGTGCATTGTCAGTGGTGGCCTTTGTCACCTGGGCCGCTGTGTCAACACAGAGGGCAGCTTCCAGTGT GTCTGCAATGCAGGCTTCGAGCTCAGCCCTGACGGCAAGAACTGTGTGGACCACAACGAGTGTGCCACCAGCACCATGTGC GTCAACGGCGTGTGTCTCAACGAGGATGGCAGCTTCTCCTGCCTCTGCAAACCCGGCTTCCTGCTGGCGCCTGGCGGCCAC TACTGCATGGACATTGACGAGTGCCAGACGCCCGGCATCTGCGTGAACGGCCACTGTACCAACACCGAGGGCTCCTTCCGC TGCCAGTGCCTGGGGGGGCTGGCGGTAGGCACGGATGGCCGCGTGTGCGTGGACACCCACGTGCGCAGCACCTGCTATGGG GCCATCGAGAAGGGCTCCTGTGCCCGCCCCTTCCCTGGCACTGTCACCAAGTCCGAGTGCTGCTGTGCCAATCCGGACCAC GGTTTTGGGGAGCCCTGCCAGCTTTGTCCTGCCAAAGACTCCGCTGAGTTCCAGGCACTGTGCAGCAGTGGGCTTGGCATT ACCACGGATGGTCGAGACATCAACGAGTGTGCTCTGGATCCTGAGGTTTGTGCCAATGGCGTGTGCGAGAACCTTCGGGGC AGCTACCGCTGTGTCTGCAACCTGGGTTATGAGGCAGGTGCCTCAGGCAAGGACTGCACAGACGTGGATGAGTGTGCCCTC AACAGCCTCCTGTGTGACAACGGGTGGTGCCAGAATAGCCCTGGCAGCTACAGCTGCTCCTGCCCCCCCGGCTTCCACTTC TGGCAGGACACGGAGATCTGCAAAGATGTCGACGAATGCCTGTCCAGCCCGTGTGTGAGTGGCGTCTGTCGGAACCTGGCC GGCTCCTACACCTGCAAATGTGGCCCTGGCAGCCGGCTGGACCCCTCTGGTACCTTCTGTCTAGACAGCACCAAGGGCACC TGCTGGCTGAAGATCCAGGAGAGCCGCTGTGAGGTGAACCTTCAGGGAGCCAGCCTGCGGTCTGAGTGCTGTGCCACCCTC GGGGCAGCCTGGGGGAGCCCCTGCGAACGCTGCGAGATCGACCCTGCCTGTGCCCGGGGCTTTGCCCGGATGACGGGTGTC ACCTGCGATGATGTGAACGAGTGTGAGTCCTTCCCGGGAGTCTGTCCCAACGGGCGTTGCGTCAACACTGCTGGGTCTTTC CGCTGTGAGTGTCCAGAGGGCCTGATGCTGGACGCCTCAGGCCGGCTGTGCGTGGATGTGAGATTGGAACCATGTTTCCTG CGATGGGATGAGGATGAGTGTGGGGTCACCCTGCCTGGCAAGTACCGGATGGACGTCTGCTGCTGCTCCATCGGGGCCGTG TGGGGAGTCGAGTGCGAGGCCTGCCCGGATCCCGAGTCTCTGGAGTTCGCCAGCCTGTGCCCGCGGGGGCTGGGCTTCGCC AGCCGGGACTTCCTGTCTGGCCGACCATTCTATAAAGATGTGAATGAATGCAAGGTGTTCCCTGGCCTCTGCACGCACGGT ACCTGCAGAAACACGGTGGGCAGCTTCCACTGCGCCTGTGCGGGGGGCTTCGCCCTGGATGCCCAGGAACGGAACTGCACA GATATCGACGAGTGTCGCATCTCTCCTGACCTCTGCGGCCAGGGCACCTGTGTCAACACGCCGGGCAGCTTTGAGTGCGAG TGTTTTCCCGGCTACGAGAGTGGCTTCATGCTGATGAAGAACTGCATGGACGTGGACGAGTGTGCAAGGGACCCGCTGCTC TGCCGGGGAGGCACTTGCACCAACACGGATGGGAGCTACAAGTGCCAGTGTCCCCCTGGGCATGAGCTGACGGCCAAGGGC ACTGCCTGTGAGGACATCGATGAGTGCTCCCTGAGTGATGGCCTGTGTCCCCATGGCCAGTGTGTCAATGTCATCGGTGCC TTCCAGTGCTCCTGCCATGCCGGCTTCCAGAGCACACCTGACCGCGGAGCTACAAGTGCCAGTTGCCCGACGGAAGGGCAT GTGCAGGTCGTCCTGGGGCCTGGGGAGCAGATGTGTACTGGGTGGTCTATCAGGGCAAAGCTAAGCACAGTCCCCAACCCT GCTCCCCCAGACGTGGACGAGTGTGAAGAGAACCCCCGCGTTTGTGACCAAGGCCACTGCACCAACATGCCAGGGGGTCAC CGCTGCCTGTGCTATGATGGCTTCATGGCCACGCCAGACATGAGGACATGTGTTGATGTGGATGAGTGTGACCTGAACCCT CACATCTGCCTCCATGGGGACTGCGAGAACACGAAGGGTTCCTTTGTCTGCCACTGTCAGCTGGGCTACATGGTCAGGAAG GGGGCCACAGGCTGCTCTGATGTGGATGAATGCGAGGTTGGAGGACACAACTGTGACAGTCACGCCTCCTGTCTCAACATC CCAGGGAGTTTCAGCTGTAGGTGCCTGCCAGGCTGGGTGGGGGATGGCTTCGAATGTCACGACCTGGATGAATGCGTCTCC CAGGAGCACCGGTGCAGCCCAAGAGGTGACTGTCTCAATGTCCCTGGCTCCTACCGCTGCACCTGCCGCCAGGGCTTTGCC GGGGATGGCTTCTTCTGCGAAGACAGGGATGAATGTGCCGAGAACGTGGACCTCTGTGACAACGGGCAGTGCCTCAATGCG CCCGGCGGGTACCGCTGTGAATGTGAGATGGGCTTTGACCCCACCGAGGACCACCGGGCCTGCCAGGATGTGGACGAGTGT GCGCAAGAGAACCTCTGTGCATTTGGGAGCTGTGAGAACCTGCCTGGAATGTTCCGCTGCATCTGCAATGGTGGCTACGAA CTGGACCGAGGGGGTGGCAACTGCACAGACATCAACGAGTGTGCAGACCCAGTAAACTGCATCAACGGCGTGTGCATTAAC ACCCCCGGCAGCTACCTCTGCAGCTGCCCCCAGGATTTTGAGCTGAACCCCAGCGGAGTGGGCTGCGTGGACACTCGGGCC GGGAACTGTTTCCTGGAGACGCATGACCGAGGGGACAGTGGCATTTCCTGCAGTGCCGAGATCGGAGTTGGTGTCACCCGA GCTTCCTGCTGTTGCTCCCTGGGCCGGGCTTGGGGCAATCCCTGTGAGCTGTGCCCTATGGCCAACACCACTGAGTACAGA ACCCTGTGCCCGGGTGGTGAGGGCTTCCAGCCTAACCGCATCACTGTCATTCTGGAAGACATCGACGAGTGCCAAGAGCTG CCAGGGCTGTGTCAGGGGGGTGACTGCGTCAACACGTTTGGCAGTTTCCAGTGTGAGTGCCCACCTGGCTACCACCTCAGT GAGCACACCCGCATCTGTGAGGATATTGACGAATGCTCCACACACTCCGGCATCTGTGGCCCTGGCACCTGCTACAACACC CTGGGGAACTACACCTGTGTCTGCCCTGCAGAGTACCTCCAAGTCAATGGTGGCAACAACTGCATGGATATGAGGAAGAGT GTCTGCTTCCGGCACTATAACGGCACATGTCAAAATGAGCTGGCCTTCAACGTGACCCGGAAAATGTGTTGCTGCTCCTAC AACATTGGCCAGGCCTGGAATAGACCCTGTGAGGCCTGCCCCACTCCCATCAGTCCTGACTACCAGATCCTGTGTGGAAAT CAGGCCCCGGGATTCCTCACTGACATCCACACGGGGAAGCCCCTTGACATTGATGAGTGTGGGGAGATCCCCGCCATCTGT GCCAATGGCATCTGCATAAACCAGATCGGGAGTTTCCGCTGCGAGTGCCCCGCAGGCTTCAACTACAACAGCATCCTGCTG GCTTGTGAAGATGTCGATGAGTGTGGCAGCAGGGAGAGTCCCTGCCAGCAGAATGCTGACTGCATCAACATCCCCGGTAGC TACCGCTGCAAGTGCACCCGAGGGTACAAACTGTCGCCAGGCGGGGCTTGTGTGGGACGGAATGAGTGTCGGGAGATCCCG AATGTCTGTAGCCATGGTGACTGCATGGACACAGAAGGCAGCTACATGTGTCTGTGTCACCGTGGATTCCAGGCCTCTGCA GACCAGACCCTGTGCATGGACATTGACGAGTGTGACCGGCAGCCTTGTGGAAATGGGACCTGCAAGAACATCATTGGCTCC TACAACTGCCTCTGCTTCCCTGGCTTTGTGGTGACACACAATGGGGATTGTGTGGATTTTGATGAGTGTACTACCCTGGTG GGGCAGGTGTGCCGATTTGGCCATTGCCTCAACACAGCTGGTTCCTTCCACTGCCTCTGCCAGGATGGCTTTGAGCTCACA GCTGATGGGAAGAACTGTGTGGACACCAATGAGTGCCTCAGCCTTGCAGGAACCTGCCTACCCGGCACTTGCCAGAACCTC GAGGGCTCCTTCCGCTGCATCTGTCCCCCTGGCTTCCAGGTGCAGAGTGACCACTGCATTGATATCGACGAGTGCTCAGAG GAGCCCAACCTCTGCCTCTTTGGCACCTGTACCAACAGCCCTGGGAGCTTCCAGTGCCTCTGCCCACCTGGCTTTGTCCTC TCTGACAATGGGCACCGTTGCTTTGACACACGGCAGAGTTTCTGCTTCACCCGTTTTGAGGCTGGGAAGTGCTCGGTGCCC AAAGCTTTCAACACCACCAAGACCCGCTGCTGCTGCAGTAAGAGGCCTGGGGAGGGCTGGGGAGACCCCTGCGAACTGTGT CCCCAGGAGGGCAGCGCTGCCTTTCAGGAGCTCTGCCCCTTTGGCCACGGGGCAGTCCCAGGCCCGGATGACTCCCGAGAA GACGTGAATGAGTGTGCAGAGAACCCTGGCGTCTGCACTAACGGCGTCTGTGTCAACACCGATGGATCCTTCCGCTGTGAG TGTCCCTTTGGCTACAGCCTGGACTTCACTGGCATCAACTGTGTGGACACAGACGAGTGCTCTGTCGGCCACCCCTGTGGG CAAGGGACATGCACCAATGTCATCGGAGGCTTCGAATGTGCCTGTGCTGACGGCTTTGAGCCTGGCCTCATGATGACCTGC GAGGAGATCGACGAATGCTCCCTGAACCCGCTGCTCTGTGCCTTCCGCTGCCACAATACCGAGGGCTCCTACCTGTGCACC TGTCCAGCCGGCTACACCCTGCGGGAGGATGGGGCCATGTGTGAAGATGTGGACGAGTGTGCAGATGGTCAGCAGGACTGC CACGCCCGGGGCATGGAGTGCAAGAACCTCATCGGTACCTTCGCGTGCGTCTGTCCCCCAGGCATGCGGCCCCTGCCTGGC TCTGGGGAGGGCTGCACAGATGACAATGAATGCCACGCTCAGCCTGACCTCTGTGTCAACGGCCGCTGTGTCAACACCGCG GGCAGCTTCCGGTGCGACTGTGATGAGGGATTCCAGCCCAGCCCCACCCTTACCGAGTGCCACGACATCCGGCAGGGGCCC TGCTTTGCCGAGGTGCTGCAGACCATGTGCCGGTCTCTGTCCAGCAGCAGTGAGGCTGTCACCAGGGCCGAGTGCTGCTGT GGGGGTGGCCGGGGCTGGGGGCCCCGCTGCGAGCTCTGTCCCCTGCCCGGCACCTCTGCCTACAGGAAGCTGTGCCCCCAT GGCTCAGGCTACACTGCTGAGGGCCGAGATGTAGATGAATGCCGTATGCTTGCTCACCTGTGTGCTCATGGGGAGTGCATC AACAGCCTTGGCTCCTTCCGCTGCCACTGTCAGGCCGGGTACACACCGGATGCTACTGCTACTACCTGCCTGGATATGGAT GAGTGCAGCCAGGTCCCCAAGCCATGTACCTTCCTCTGCAAAAACACGAAGGGCAGTTTCCTGTGCAGCTGTCCCCGAGGC TACCTGCTGGAGGAGGATGGCAGGACCTGCAAAGACCTGGACGAATGCACCTCCCGGCAGCACAACTGTCAGTTCCTCTGT GTCAACACTGTGGGCGCCTTCACCTGCCGCTGTCCGCCCGGCTTCACCCAGCACCACCAGGCCTGCTTCGACAATGATGAG TGCTCAGCCCAGCCTGGCCCATGTGGTGCCCACGGGCACTGCCACAACACCCCGGGCAGCTTCCGCTGTGAATGCCACCAA GGCTTCACCCTGGTCAGCTCAGGCCATGGCTGTGAAGATGTGAATGAATGTGATGGGCCCCACCGCTGCCAGCATGGCTGT CAGAACCAGCTAGGGGGCTACCGCTGCAGCTGCCCCCAGGGTTTCACCCAGCACTCCCAGTGGGCCCAGTGTGTGGATGAG AATGAGTGTGCCCTGTCGCCCCCCACCTGCGGGAGCGCCTCCTGTCGCAACACTCTTGGTGGCTTCCGCTGCGTCTGCCCC TCTGGCTTTGACTTTGATCAGGCCCTCGGGGGCTGCCAGGAGGTGGATGAGTGCGCCGGACGGCGTGGCCCCTGTAGCTAC AGCTGTGCCAACACGCCTGGTGGCTTCCTGTGCGGCTGTCCTCAAGGCTACTTCCGGGCTGGGCAAGGGCACTGTGTCTCC GGCCTGGGCTTCAGCCCCGGACCCCAGGACACCCCGGACAAAGAGGAGCTGCTCTCGTCTGAAGCCTGCTACGAATGCAAG ATCAATGGCCTCTCCCCTCGGGACCGGCCACGACGCAGTGCCCACAGGGACCACCAGGTGAACCTGGCCACCCTTGACTCC GAGGCCCTGCTGACCTTGGGCCTGAACCTCTCACACCTGGGCCGGGCCGAGCGCATCCTGGAGCTCCGGCCGGCCCTGGAG GGTCTAGAGGGCCGGATCCGCTACGTCATCGTCCGCGGAAACGAGCAAGGTTTCTTTCGCATGCATCACCTCCGTGGCGTC AGCTCCCCGCAGCTGGGGCGGAGGCGGCCGGGGCCTGGAACCTACCGGCTGGAGGTGGTGAGCCACATGGCAGGACCCTGG GGTGTCCAGCAAGAGGGGCAGCCAGGGCCATGGGGCCAGGCCTTGAGGCTGAAGGTGCAACTGTCAGTTGCTTTAGTTGGG AGGAGCCTCAGTGGGCCCCAGCTGTCCAGAGAAGGGGGATTCTGGAACTGGGAAGGACTGATCCCCAGAAGCGATGGCTGA CCAGATTGAACCCCGAAACTCAGGAAGAGTGAAATGCTACACGACAACCTCAGGCAAGCCCGGCCTCTGCCTGGGCCTCTG TGCCAGCCCCGGGGGCCCCCCAGTTACTCAGTCTTTCCTGGAGACAGCAAGAAGCTGCAATGTGCAATCCCCCTGCCCCCA CAGCCAAGGTCAGGAAGAGGCCCTGTGGTCACCGTGTCTGGCCAATCTCAGGCTTTCACTTCTGTACTGCACTGTGGCTTG CCCTGGCGGGGGGCAGGGGGTTGGCAGGACATGGCAATGGGCAACTGGGGTGGGCACAGGGCTTATTCCTCGGAGTAGAAG GGTGTACAGGGGGCCCAGACTCCACAGTGACTTGCCACATTTGCCCCCCATTTGGAGAATGCTTTTATATCAAAAGTGGAG ACGATAATAAAGTTATTTTGGGTTAAGTCTGCCTGCCCTTTGGCAAGTTCTTGAAGTAAGTAGATGCTGCCCTCGGACTGG GCGAGGCAGATCTTGTGCCTGGGGAAGCAGAAGGCCTTATGGGCTCCCCAGAATGGTAATAATGGCTCACGCTTCCTGACC ACGTACTACATACCAGACACCATTCGATTTTTTTTTTATTTTTTCTGAGACAGGGTCTTGCTCTGTTGCCTAGGTTGGAGT GCAGTGGCGCGATCGTGGCTCCCTGTGGTCGCCACTTTCCGGGCTCGAGCAATCCTCCCACCTCAGCCTCTCCCAAGTAGC TGGGACCAAAGGTGCACGCCACCACACCCAGCTAATTTTTTTAAATTTCTTTTTCTTTCTCTTTCTTTTTGAGACCAGCCC GGCCAACATGACGAAACCCCATCTCTAATA

The disclosed NOV2a nucleic acid sequence, localized to chromsome 19, has 3196 of 4382 bases (72%) identical to a Mus musculus fibrillin 2 (fbn2) mRNA (gb:GENBANK- ID:MUSFBN2|acc:L39790) (E = 0.0).

A NOV2a polypeptide (SEQ ID NO:6) encoded by SEQ ID NO:5 has 2995 amino acid residues and is presented using the one-letter code in Table 2B. Signal P, Psort and/or Hydropathy results predict that NOV2a contains a signal peptide and is likely to be localized to the endoplasmic reticulum membrane with a certainty of 0.5500. The most likely cleavage site for a NOV2a peptide is between amino acids 54 and 55, at: TSS-RK.

Table 2B. Encoded NOV2a protein sequence (SEQ ID NO:6).

MSKSCMT LCPLLQRLAPPGPTWPS PSLPLPTPVLPWLCT SPKTLLXHHTSSRKPPLTVHRFPVGAGCLLG PEAPGLP HLATTLPSS CWVPHGXGGGLIXGXLCF SAASRGHAMTLEGLYLARGPLARLLLAWSAL CMAGGQGRWDGALEAAGPGR VRRRGSPGI QGPNVCGSRFHAYCCPG RTFPGRSQCVVPICRRACGEGFCSQPNLCTCADGT APSCGVSRGSGCSVSCM NGGTCRGASCLCQKGYTGTVCGQPICDRGCHNGGRCIGPNRCACVYGFMGPQCERDYRTGPCFGQVGPEGCQHQLTGLVCT KALCCATVGRA GLPCELCPAQPHPCRRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVGHRLSDSSAAC EDYDECSTIPGICEGGECTNTVSSYFCKCPPGFYTSPDGTLHGQSRAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWA AGPVPELCPPRGSNEFQQLCAQRLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARGIPSLGPGNSNIGTATLNQ TIDICRHFTN CLNGRCLPTPSSYRCECNVGYTQDVRGECIDVDECTSSPCHHGDCVNIPGTYHCRCYPGFQATPTRQACV DVDECIVSGG CHLGRCVNTEGSFQCVCNAGFELSPDGKNCVDHNECATSTMCVNGVCLNEDGSFSCLCKPGFLLAPGGHY CMDIDECQTPGICVNGHCTNTEGSFRCQC GGLAVGTDGRVCVDTHVRSTCYGAIEKGSCARPFPGTVTKSECCCANPDHG FGEPCQLCPAKDSAEFQALCSSGLGITTDGRDINECA DPEVCANGVCENLRGSYRCVCN GYEAGASGKDCTDVDECA N SL CDNGWCQNΞPGSYSCSCPPGFHFWQDTEICKDVDEC SSPCVSGVCRN AGSYTCKCGPGSRLDPSGTFCLDSTKGTC LKIQESRCEVNLQGASLRSECCATLGAAWGSPCERCEIDPACARGFARMTGVTCDDVNECESFPGVCPNGRCVNTAGSFR CECPEGLMLDASGRLCVDVRLEPCFLRWDEDECGVTLPGKYRMDVCCCSIGAVWGVECEACPDPESLEFASLCPRGLGFAS RDF SGRPFYKDVNECKVFPGLCTHGTCRNTVGSFHCACAGGFALDAQERNCTDIDECRISPDLCGQGTCVNTPGSFECEC FPGYESGFMLMKNCMDVDECARDPLLCRGGTCTNTDGSYKCQCPPGHELTAKGTACEDIDECSLSDG CPHGQCVNVIGAF QCSCHAGFQSTPDRGATSASCPTEGHVQVV GPGEQMCTG SIRAK STVPNPAPPDVDECEENPRVCDQGHCTNMPGGHR CLCYDGFMATPDMRTCVDVDECDLNPHIC HGDCENTKGSFVCHCQ GYMVRKGATGCSDVDECEVGGHNCDSHASCLNIP GSFΞCRCLPGWVGDGFECHD DECVSQEHRCSPRGDC NVPGSYRCTCRQGFAGDGFFCEDRDECAENVDLCDNGQC NAP GGYRCECEMGFDPTEDHRACQDVDECAQENLCAFGSCENLPGMFRCICNGGYELDRGGGNCTDINECADPVNCINGVCINT PGSYLCSCPQDFELNPSGVGCVDTRAGNCFLETHDRGDSGISCSAEIGVGVTRASCCCSLGRAWGNPCE CPMANTTEYRT LCPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCECPPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTL GNYTCVCPAEYLQVNGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCEACPTPISPDYQILCGNQ APGFLTDIHTGKPLDIDECGEIPAICANGICINQIGSFRCECPAGFNYNSILLACEDVDECGSRESPCQQNADCINIPGSY RCKCTRGYKLSPGGACVGRNECREIPNVCSHGDCMDTEGSYMCLCHRGFQASADQTLCMDIDECDRQPCGNGTCKNIIGSY NCLCFPGFVVTHNGDCVDFDECTTLVGQVCRFGHC NTAGSFHCLCQDGFELTADGKNCVDTNECLSLAGTC PGTCQNLE GSFRCICPPGFQVQSDHCIDIDECSEEPNLC FGTCTNSPGSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPK AFNTTKTRCCCSKRPGEG GDPCELCPQEGSAAFQELCPFGHGAVPGPDDSREDVNECAENPGVCTNGVCVNTDGΞFRCEC PFGYSLDFTGINCVDTDECSVGHPCGQGTCTNVIGGFECACADGFEPGLMMTCEEIDECSLNPLLCAFRCHNTEGSYLCTC PAGYTLREDGAMCEDVDECADGQQDCHARGMECKNLIGTFACVCPPGMRPLPGSGEGCTDDNECHAQPD CVNGRCVNTAG SFRCDCDEGFQPSPT TECHDIRQGPCFAEVLQTMCRSLSSSSEAVTRAECCCGGGRG GPRCE CP PGTSAYRK CPHG SGYTAEGRDVDECRMLAHLCAHGECINSLGSFRCHCQAGYTPDATATTCLDMDECSQVPKPCTF CKNTKGSF CSCPRGY LLEEDGRTCKDLDECTSRQHNCQF CVNTVGAFTCRCPPGFTQHHQACFDNDECSAQPGPCGAHGHCHNTPGSFRCECHQG FTLVSSGHGCEDVNECDGPHRCQHGCQNQ GGYRCSCPQGFTQHSQ AQCVDENECA SPPTCGSASCRNTLGGFRCVCPS GFDFDQALGGCQEVDECAGRRGPCSYSCANTPGGFLCGCPQGYFRAGQGHCVSG GFSPGPQDTPDKEELLSSEACYECKI NGLSPRDRPRRSAHRDHQVN AT DSEAL TLGLN SHLGRAERI ELRPA EGLEGRIRYVIVRGNEQGFFRMHHLRGVS SPQLGRRRPGPGTYRLEVVSHMAGP GVQQEGQPGPWGQA RLKVQ SVALVGRΞLSGPQLSREGGF NWEG IPRSDG

The NOV2a amino acid sequence has 1895 of 2810 amino acid residues (67%) identical to, and 2257 of 2810 amino acid residues (80%) similar to, a Mus musculus 2907 amino acid residue fibrillin 2 precursor protein (ptnr:SWISSPROT-ACC:Q61555) (E = 0.0).

The disclosed NOV2a is expressed in at least the following tissues: testes, ovary, lung, liver, B-cells, total-fetus, spleen, Nervous System, Brain, Prosencephalon/Forebrain, Diencephalon, Pituitary Gland, Hematopoietic Tissues, Lymphoid tissue, Lymph node and Whole Organism. This information was derived by determining the tissue sources ofthe sequences that were included in the invention. In addition, the NOV2a sequence is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus fibrillin 2 (fbn2) gene (GENBANK-ID: gb:GENBANK- ID:MUSFBN2|acc:L39790): testes, ovary, lung, liver, total-fetus, brain, spleen, Nervous System, Brain, Pituitary Gland; Hematopoietic and Lymphatic System.

NOV2b A disclosed NOV2b nucleic acid of 9894 nucleotides (also referred to as 153568997) encoding a novel Fibrillin-like protein is shown in Table 2C. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 520-522 and ending with a TGA codon at nucleotides 9052-9054. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table 2C, and the start and stop codons are in bold letters. Table 2C. NOV2b nucleotide sequence (SEQ ID NO:7).

CCTCCC_ATTCCTCTCCTCTCTC_TCCCATCTGGAC_AGCCCCCAGCCTTCTGACACCCTGTTTCCCCCCCGGGACCCTGCTGC CCTTCCCC_ACACC_ACCT_AAC_ATGC_ATTTCTGACCTTGCTCCTCCCACTCAAAGCTTTTCAAGGGCTCTTCATTGTCCTTGΆ CG_AATG_AGC_AAAAGCTGCATG_AC_ACTGCTATGCCCΆCTCCTGCAACGCCTGGCCCCACCTGGCCCCACCTGGCCCTCTCTC CCCTCCCTTCC_ACTTCCC_ACTCCCGTCTTGCCTTGGCTGTGCACCCTCAGCCCCAAGACCCTTCTCTAACATCACACCTCC TCCAGGAAGCCTCCTCTG_ACTGTTCACCGCTTCCCGGTTGGGGCTGGATGCCTCCTTGGGCTCCCAGAGGCCCCGGGGCTG CCCC_ATTTGGCA_ACC_ACCCTGCCCTCCTCGTTGTGTTGGGTCCCCCATGGCTAGGGTGGGGGGCTCATCTAGGGCTGACTC TGCTTCCTCTCCGCAGCCTCCAGGGGACACGCCATGACTCTGGAGGGTCTGTATTTGGCAAGGGGCCCCCTGGCCCGGCTC CTGCTGGCCTG_GTCGGCCCTGTTGTGCATGGCAGGTGGCCAAGGCCGCTGGGACGGGGCCTTGGAGGCTGCAGGTCCTGGA CGTGTGCGGAGGCGGGGC_AGCCCAGGCATCTTGCAGGGGCCGAATGTGTGCGGCTCCCGGTTCCATGCCTACTGCTGTCCA GGCTGGAGG_ACATTCCCTGGCAGGAGCCAGTGTGTCGTACCCATCTGTAGGCGCGCCTGCGGTGAAGGCTTCTGCTCCCAG CCC_AACCTGTGC_ACCTGTGCGG_ATGGGACGCTGGCTCCCAGCTGCGGGGTGAGCCGAGGGTCAGGGTGCAGTGTGAGCTGT _ATGAATGGGGGCACCTGCCGGGGGGCGTCCTGTCTGTGTCAGAAGGGCTACACAGGCACCGTGTGTGGGCAGCCCATCTGT G_ACCGCGGCTGCCAC_AATGGGGGTCGCTGCATTGGGCCCAACCGCTGCGCCTGTGTGTATGGCTTCATGGGACCTCAATGT G_AGAGAG_ATT_ACCGGACGGGATCCTGCTTTGGCCAAGTAGGCCCCGAGGGGTGCCAGCATCAGCTGACGGGCCTCGTGTGC ACCAAGGCACTTTGCTGTGCCACTGTGGGCCGTGCCTGGGGCCTTCCATGTGAACTTTGCCCTGCACAGCCACACCCCTGC CGCCGCGGCTTCATCCCCAATATCCACACGGGGGCCTGCCAAGATGTGGATGAGTGCCAGGCTGTGCCAGGCCTGTGCCAG GGAGGCAGCTGCGTCAACATGGTGGGCTCCTTCCATTGCCGCTGTCCAGTTGGACACCGGCTCAGTGACAGCAGCGCCGCA TGTGAAGACTACCGGGCCGGCGCCTGCTTCTCAGTGCTTTTCGGGGGCCGCTGTGCTGGAGACCTCGCCGGCCACTACACT CGCAGGCAGTGCTGCTGTGACAGGGGCAGGTGCTGGGCAGCTGGCCCGGTCCCTGAGCTGTGTCCTCCTCGGGGCTCCAAT GAATTCCAGCAACTGTGCGCCCAGCGGCTGCCGCTGCTACCCGGCCACCCTGGCCTCTTCCCTGGCCTCCTGGGCTTCGGA TCCAATGGCATGGGTCCCCCTCTTGGGCCAGCGCGACTCAACCCCCATGGCTCTGATGCGCGTGGGATCCCCAGCCTGGGC CCTGGCAACTCTAATATTGGCACTGCTACCCTGAACCAGACCATTGACATCTGCCGACACTTCACCAACCTGTGTCTGAAT GGCCGCTGCCTGCCCACGCCTTCCAGCTACCGCTGCGAGTGTAACGTGGGCTACACCCAGGACGTGCGCGGCGAGTGCATT GATGTAGACGAATGCACCAGCAGCCCCTGCCACCACGGTGACTGCGTCAACATCCCCGGCACCTACCACTGCCGGTGCTAC CCGGGCTTCCAGGCCACGCCCACCAGGCAGGCATGCGTGGATGTGGACGAGTGCATTGTCAGTGGTGGCCTTTGTCACCTG GGCCGCTGTGTCAACACAGAGGGCAGCTTCCAGTGTGTCTGCAATGCAGGCTTCGAGCTCAGCCCTGACGGCAAGAACTGT GTGGACCACAACGAGTGTGCCACCAGCACCATGTGCGTCAACGGCGTGTGTCTCAACGAGGATGGCAGCTTCTCCTGCCTC TGCAAACCCGGCTTCCTGCTGGCGCCTGGCGGCCACTACTGCATGGACATTGACGAGTGCCAGACGCCCGGCATCTGCGTG AACGGCCACTGTACCAACACCGAGGGCTCCTTCCGCTGCCAGTGCCTGGGGGGGCTGGCGGTAGGCACGGATGGCCGCGTG TGCGTGGACACCCACGTGCGCAGCACCTGCTATGGGGCCATCGAGAAGGGCTCCTGTGCCCGCCCCTTCCCTGGCACTGTC ACCAAGTCGGAGTGCTGCTGTGCCAATCCGGACCACGGTTTTGGGGAGCCCTGCCAGCTTTGTCCTGCCAAAAACTCCGCT GAGTTCCAGGCACTGTGCAGCAGTGGGCTTGGCATTACCACGGATGGTCGAGACATCAACGAGTGTGCTCTGGATCCTGAG GTTTGTGCCAATGGCGTGTGCGAGAACCTTCGGGGCAGCTACCGCTGTGTCTGCAACCTGGGTTATGAGGCAGGTGCCTCA GGCAAGGACTGCACAGACGTGGATGAGTGTGCCCTCAACAGCCTCCTGTGTGACAACGGGTGGTGCCAGAATAGCCCTGGC AGCTACAGCTGCTCCTGCCCCCCCGGCTTCCACTTCTGGCAGGACACGGAGATCTGCAAAGATGTCGACGAATGCCTGTCC AGCCCGTGTGTGAGTGGCGTTTGTCGGAACCTGGCCGGCTCCTACACCTGCAAATGTGGCCCTGGCAGCCGGCTGGACCCC TCTGGTACCTTCTGTCTAGACAGCACCAAGGGCACCTGCTGGCTGAAGATCCAGGAGAGCCGCTGTGAGGTGAACCTTCAG GGAGCCAGCCTGCGGTCTGAGTGCTGTGCCACCCTCGGGGCAGCCTGGGGGAGCCCCTGCGAACGCTGCGAGATCGACCCT GCCTGTGCCCGGGGCTTTGCCCGGATGACGGGTGTCACCTGCGATGATGTGAACGAGTGTGAGTCCTTCCCGGGAGTCTGT CCCAACGGGCGTTGCGTCAACACTGCTGGGTCTTTCCGCTGTGAGTGTCCAGAGGGCCTGATGCTGGACGCCTCAGGCCGG CTGTGCGTGGATGTGAGATTGGAACCATGTTTCCTGCGATGGGATGAGGATGAGTGTGGGGTCACCCTGCCTGGCAAGTAC CGGATGGACGTCTGCTGCTGCTCCATCGGGGCCGTGTGGGGAGTCGAGTGCGAGGCCTGCCCGGATCCCGAGTCTCTGGAG TCGCCAGCCTGTGCCCGCGGGGGCTGGGCTTCGCCAGCCGGGACTTCCTGTCTGGCCGACCATTCTATAAAGATGTGAAT GAATGCAAGGTGTTCCCTGGCCTCTGCACGCACGGTACCTGCAGAAACACGGTGGGCAGCTTCCACTGCGCCTGTGCGGGG GGCTTCGCCCTGGATGCCCAGGAACGGAACTGCACAGATATCGACGAGTGTCGCATCTCTCCTGACCTCTGCGGCCAGGGC ACCTGTGTCAACACGCCGGGCAGCTTTGAGTGCGAGTGTTTTCCCGGCTACGAGAGTGGCTTCATGCTGATGAAGAACTGC ATGGACGTGGACGAGTGTGCAAGGGACCCGCTGCTCTGCCGGGGAGGCACTTGCACCAACACGGATGGGAGCTACAAGTGC CAGTGTCCCCCTGGGCATGAGCTGACGGCCAAGGGCACTGCCTGTGAGGACATCGATGAGTGCTCCCTGAGTGATGGCCTG TGTCCCCATGGCCAGTGTGTCAATGTCATCGGTGCCTTCCAGTGCTCCTGCCATGCCGGCTTCCAGAGCACACCTGACCGC CAGGGCTGCGTGGACATCAACGAATGCCGGGTCCAGAATGGTGGGTGTGACGTGCACCGTATTAACACTGAGGGCAGCTAC CGGTGCAGCTGTGGGCAGGGCTACTCGCTGATGCCCGACGGAAGGGCATGTGCAGACGTGGACGAGTGTGAAGAGAACCCC CGCGTTTGTGACCAAGGCCACTGCACCAACATGCCAGGGGGTCACCGCTGCCTGTGCTATGATGGCTTCATGGCCACGCCA GACATGAGGACATGTGTTGATGTGGATGAGTGTGACCTGAACCCTCACATCTGCCTCCATGGGGACTGCGAGAACACGAAG GGTTCCTTTGTCTGCCACTGTCAGCTGGGCTACATGGTCAGGAAGGGGGCCACAGGCTGCTCTGATGTGGATGAATGCGAG GTTGGAGGACACAACTGTGACAGTCACGCCTCCTGTCTCAACATCCCGGGGAGTTTCAGCTGTAGGTGCCTGCCAGGCTGG GTGGGGGATGGCTTCGAATGTCACGACCTGGATGAATGCGTCTCCCAGGAGCACCGGTGCAGCCCAAGAGGTGACTGTCTC AATGTCCCTGGCTCCTACCGCTGCACCTGCCGCCAGGGCTTTGCCGGGGATGGCTTCTTCTGCGAAGACAGGGATGAATGT GCCGAGAACGTGGACCTCTGTGACAACGGGCAGTGCCTCAATGCGCCCGGCGGGTACCGCTGTGAATGTGAGATGGGCTTT GACCCCACCGAGGACCACCGGGCCTGCCAGGATGTGGACGAGTGTGCGCAAGAGAACCTCTGTGCATTTGGGAGCTGTGAG AACCTGCCTGGAATGTTCCGCTGCATCTGCAATGGTGGCTACGAACTGGACCGAGGGGGTGGCAACTGCACAGACATCAAC GAGTGTGCAGACCCAGTAAACTGCATCAACGGCGTGTGCATTAACACCCCCGGCAGCTACCTCTGCAGCTGCCCCCAGGAT TTTGAGCTGAACCCCAGCGGAGTGGGCTGCGTGGACACTCGGGCCGGGAACTGTTTCCTGGAGACGCATGACCGAGGGGAC AGTGGCATTTCCTGCAGTGCCGAGATCGGAGTTGGTGTCACCCGAGCTTCCTGCTGTTGCTCCCTGGGCCGGGCTTGGGGC AATCCCTGTGAGCTGTGCCCTATGGCCAACACCACTGAGTACAGAACCCTGTGCCCGGGTGGTGAGGGCTTCCAGCCTAAC CGCATCACTGTCATTCTGGAAGACATCGACGAGTGCCAAGAGCTGCCAGGGCTGTGTCAGGGGGGTGACTGCGTCAACACG TTTGGCAGTTTCCAGTGTGAGTGCCCACCTGGCTACCACCTCAGTGAGCACACCCGCATCTGTGAGGATATTGACGAATGC TCCACACACTCCGGCATCTGTGGCCCTGGCACCTGCTACAACACCCTGGGGAACTACACCTGTGTCTGCCCTGCAGAGTAC CTCCAAGTCAATGGTGGCAACAACTGCATGGATATGAGGAAGAGTGTCTGCTTCCGGCACTATAACGGCACATGTCAAAAT GAGCTGGCCTTCAACGTGACCCGGAAAATGTGTTGCTGCTCCTACAACATTGGCCAGGCCTGGAATAGACCCTGTGAGGCC TGCCCCACTCCCATCAGTCCTGACTACCAGATCCTGTGTGGAAATCAGGCCCCGGGATTCCTCACTGACATCCACACGGGG AAGCCCCTTGACATTGATGAGTGTGGGGAGATCCCCGCCATCTGTGCCAATGGCATCTGCATAAACCAGATCGGGAGTTTC CGCTGCGAGTGCCCCGCAGGCTTCAACTACAACAGCATCCTGCTGGCTTGTGAAGATGTCGATGAGTGTGGCAGCAGGGAG AGTCCCTGCCAGCAGAATGCTGACTGCATCAACATCCCCGGTAGCTACCGCTGCAAGTGCACCCGAGGGTACAAACTGTCG CCAGGCGGGGCTTGTGTGGGACGGAATGAGTGTCGGGAGATCCCGAATGTCTGTAGCCATGGTGACTGCATGGACACAGAA GGCAGCTACATGTGTCTGTGTCACCGTGGATTCCAGGCCTCTGCAGACCAGACCCTGTGCATGGACATTGACGAGTGTGAC CGGCAGCCTTGTGGAAATGGGACCTGCAAGAACATCATTGGCTCCTACAACTGCCTCTGCTTCCCTGGCTTTGTGGTGACA CACAATGGGGATTGTGTGGATTTTGATGAGTGTACTACCCTGGTGGGGCAGGTGTGCCGATTTGGCCATTGCCTCAACACA GCTGGTTCCTTCCACTGCCTCTGCCAGGATGGCTTTGAGCTCACAGCTGATGGGAAGAACTGTGTGGACACCAATGAGTGC CTCAGCCTTGCAGGAACCTGCCTACCCGGCACTTGCCAGAACCTCGAGGGCTCCTTCCGCTGCATCTGTCCCCCTGGCTTC CAGGTGCAGAGTGACCACTGCATTGATATCGACGAGTGCTCAGAGGAGCCCAACCTCTGCCTCTTTGGCACCTGTACCAAC AGCCCTGGGAGCTTCCAGTGCCTCTGCCCACCTGGCTTTGTCCTCTCTGACAATGGGCACCGTTGCTTTGACACACGGCAG AGTTTCTGCTTCACCCGTTTTGAGGCTGGGAAGTGCTCGGTGCCCAAAGCTTTCAACACCACCAAGACCCGCTGCTGCTGC AGTAAGAGGCCTGGGGAGGGCTGGGGAGACCCCTGCGAACTGTGTCCCCAGGAGGACTCCCCTCCCCCTCTCCGTCCAGCT GCCTTTCAGGAGCTCTGCCCCTTTGGCCACGGGGCAGTCCCAGGCCCGGATGACTCCCGAGAAGACGTGAATGAGTGTGCA GAGAACCCTGGCGTCTGCACTAACGGCGTCTGTGTCAACACCGATGGATCCTTCCGCTGTGAGTGTCCCTTTGGCTACAGC CTGGACTTCACTGGCATCAACTGTGAGGACACAGACGAGTGCTCTGTCGGCCACCCCTGTGGGCAAGGGACATGCACCAAT GTCATCGGAGGCTTCGAATGTGCCTGTGCTGACGGCTTTGAGCCTGGCCTCATGATGACCTGCGAGGACATCGACGAATGC TCCCTGAACCCGCTGCTCTGTGCCTTCCGCTGCCACAATACCGAGGGCTCCTACCTGTGCACCTGTCCAGCCGGCTACACC CTGCGGGAGGATGGGGCCATGTGTCGAGATGTGGACGAGTGTGCAGATGGTCAGCAGGACTGCCACGCCCGGGGCATGGAG TGCAAGAACCTCATCGGTACCTTCGCGTGCGTCTGTCCCCCAGGCATGCGGCCCCTGCCTGGCTCTGGGGAGGGCTGCACA GATGACAATGAATGCCACGCTCAGCCTGACCTCTGTGTCAACGGCCGCTGTGTCAACACCGCGGGCAGCTTCCGGTGCGAC TGTGATGAGGGATTCCAGCCCAGCCCCACCCTTACCGAGTGCCACGACATCCGGCAGGGGCCCTGCTTTGCCGAGGTGCTG CAGACCATGTGCCGGTCTCTGTCCAGCAGCAGTGAGGCTGTCACCAGGGCCGAGTGCTGCTGTGGGGGTGGCCGGGGCTGG GGGCCCCGCTGCGAGCTCTGTCCCCTGCCCGGCACCTCTGCCTACAGGAAGCTGTGCCCCCATGGCTCAGGCTACACTGCT GAGGGCCGAGATGTAGATGAATGCCGTATGCTTGCTCACCTGTGTGCTCATGGGGAGTGCATCAACAGCCTTGGCTCCTTC CGCTGCCACTGTCAGGCCGGGTACACACCGGATGCTACTGCTACTACCTGCCTGGATATGGATGAGTGCAGCCAGGTCCCC AAGCCATGTACCTTCCTCTGCAAAAACACGAAGGGCAGTTTCCTGTGCAGCTGTCCCCGAGGCTACCTGCTGGAGGAGGAT GGCAGGACCTGCAAAGACCTGGACGAATGCACCTCCCGGCAGCACAACTGTCAGTTCCTCTGTGTCAACACTGTGGGCGCC TTCACCTGCCGCTGTCCACCCGGCTTCACCCAGCACCACCAGGCCTGCTTCGACAATGATGAGTGCTCAGCCCAGCCTGGC CCATGTGGTGCCCACGGGCACTGCCACAACACCCCGGGCAGCTTCCGCTGTGAATGCCACCAAGGCTTCACCCTGGTCAGC TCAGGCCATGGCTGTGAAGATGTGAATGAATGTGATGGGCCCCACCGCTGCCAGCATGGCTGTCAGAACCAGCTAGGGGGC TACCGCTGCAGCTGCCCCCAGGCTTTCACCCAGCACTCCCAGTGGGCCCAGTGTGTGGATGAGAATGAGTGTGCCCTGTCG CCCCCCACCTGCGGGAGCGCCTCCTGTCGCAACACTCTTGGTGGCTTCCGCTGCGTCTGCCCCTCCGGCTTTGACTTTGAT CAGGCCCTCGGGGGCTGCCAGGAGGTGGATGAGTGCGCCGGACGGCGTGGCCCCTGTAGCTACAGCTGTGCCAACACGCCT GGTGGCTTCCTGTGCGGCTGTCCTCAAGGCTACTTCCGGGTTGGGCAAGGGCACTGTGTCTCCGGCCTGGGCTTCAGCCCC GGACCCCAGGACACCCCGGACAAAGAGGAGCTGCTCTCGTCTGAAGCCTGCTACGAATGCAAGATCAATGGCCTCTCCCCT CGGGACCGGCCACGACGCAGTGCCCACAGGGACCACCAGGTGAACCTGGCCACCCTTGACTCCGAGGCCCTGCTGACCTTG GGCCTGAACCTCTCACACCTGGGCCGGGCCGAGCGCATCCTGGAGCTCCGGCCGGCCCTGGAGGGTCTAGAGGGCCGGATC CGCTACGTCATCGTCCGCGGAAACGAGCAAGGTTTCTTTCGCATGCATCACCTCCGTGGCGTCAGCTCCCTGCAGCTGGGG CGGAGGCGGCCGGGGCCTGGAACCTACCGGCTGGAGGTGGTGAGCCACATGGCAGGACCCTGGGGTGTCCAGCAAGAGGGG CAGCCAGGGCCATGGGGCCAGGCCTTGAGGCTGAAGGTGCAACTGTCAGTTGCTTTAGTTGGGAGGAGCCTCAGTGGGCCC CAGCTGTCCAGAGAAGGGGGATTCTGGAACTGGGAAGGACTGATCCCCAGAAGCGATGGCTGACCAGATTGAACCCCGAAA CTCAGGAAGAGTGAAATGCTACACGACAACCTCAGGCAAGCCCGGCCTCTGCCTGGGCCTCTGTGCCAGCCCCGGGGGCCC CCCAGTTACTCAGTCTTTCCTGGAGACAGCAAGAAGCTGCAATGTGCAATCCCCCTGCCCCCACAGCCAAGGTCAGGAAGA GGCCCTGTGGTCACCGTGTCTGGCCAATCTCAGGCTTTCACTTCTGTACTGCACTGTGGCTTGCCCTGGCGGGGGGCAGGG GGTTGGCAGGACATGGCAATGGGCAACTGGGGTGGGCACAGGGCTTATTCCTCGGAGTAGAAGGGTGTACAGGGGGCCCAG ACTCCACAGTGACTTGCCACATTTGCCCCCCATTTGGAGAATGCTTTTATATCAAAAGTGGAGACGATAATAAAGTTATTT TGGGTTAAGTCTGCCTGCCCTTTGGCAAGTTCTTGAAGTAAGTAGATGCTGCCCTCGGACTGGGCGAGGCAGATCTTGTGC CTGGGGAAGCAGAAGGCCTTATGGGCTCCCCAGAATGGTAATAATGGCTCACGCTTCCTGACCACGTACTACATACCAGAC ACCATTCGATTTTTTTTTTATTTTTTCTGAGACAGGGTCTTGCTCTGTTGCCTAGGTTGGAGTGCAGTGGCGCGATCGTGG CTCCCTGTGGTCGCCACTTTCCGGGCTCGAGCAATCCTCCCACCTCAGCCTCTCCCAAGTAGCTGGGACCAAAGGTGCACG CCACCACACCCAGCTAATTTTTTTAAATTTCTTTTTCTTTCTCTTTCTTTTTGAGACCAGCCCGGCCAACATGACGAAACC CCATCTCTAATA

The disclosed NOV2b nucleic acid sequence, localized to chromsome 19, has 3617 of 5008 bases (72%) identical to a Rattus norvegicus fibrillin 2 (fbn2) mRNA (gb:GENBANK- ID:AF135060|acc:AF135060.1) (E = 0.0).

A NOV2b polypeptide (SEQ ID NO:8) encoded by SEQ ID NO:7 has 2844 amino acid residues and is presented using the one-letter code in Table 2D. Signal P, Psort and/or Hydropathy results predict that NOV2b contains a signal peptide and is likely to be localized to the nucleus with a certainty of 0.6000. Although PSORT suggests that the Fibrillin-like protein may be localized in the nucleus, the NOV2b protein is similar to the Fibrillin family, some members of which are released extracellularly. Therefore it is likely that NOV2b protein shows a similar localization. The most likely cleavage site for a NOV2b peptide is between amino acids 29 and 30, at: AGG-QG. Table 2D. Encoded NOV2b protein sequence (SEQ ID NO:8). TLEGLY ARGPLAR LLAWSA LCMAGGQGRWDGALEAAGPGRVRRRGSPGILQGPNVCGSRFHAYCCPGWRTFPGRSQC VVPICRRACGEGFCSQPNLCTCADGTLAPSCGVSRGSGCSVSCMNGGTCRGASCLCQKGYTGTVCGQPICDRGCHNGGRCI GPNRCACVYGFMGPQCERDYRTGSCFGQVGPEGCQHQLTGLVCTKALCCATVGRAWGLPCELCPAQPHPCRRGFIPNIHTG ACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVGHRLSDSSAACEDYRAGACFSV FGGRCAGDLAGHYTRRQCCCDRGRC WAAGPVPELCPPRGSNEFQQ CAQR PL PGHPG FPGLLGFGSNGMGPPLGPARLNPHGSDARGIPSLGPGNSNIGTAT NQTIDICRHFTNLCLNGRCLPTPSSYRCECNVGYTQDVRGECIDVDECTSSPCHHGDCVNIPGTYHCRCYPGFQATPTRQA CVDVDECIVSGGLCH GRCVNTEGSFQCVCNAGFELSPDGKNCVDHNECATSTMCVNGVC NEDGSFSCLCKPGFLLAPGG HYCMDIDECQTPGICVNGHCTNTEGSFRCQCLGGLAVGTDGRVCVDTHVRSTCYGAIEKGSCARPFPGTVTKSECCCANPD HGFGEPCQLCPAKNSAEFQALCSSGLGITTDGRDINECALDPEVCANGVCENLRGSYRCVCNLGYEAGASGKDCTDVDECA NSL CDNG CQNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRN AGSYTCKCGPGSRLDPSGTFCLDSTKG TC KIQESRCEVN QGAS RSECCATLGAAWGSPCERCEIDPACARGFARMTGVTCDDVNECESFPGVCPNGRCVNTAGS FRCECPEG MLDASGRLCVDVR EPCF RWDEDECGVTLPGKYRMDVCCCSIGAV GVECEACPDPESLEFAS CPRG GF ASRDFLSGRPFYKDVNECKVFPGLCTHGTCRNTVGSFHCACAGGFALDAQERNCTDIDECRISPDLCGQGTCVNTPGSFEC ECFPGYESGFMLMKNCMDVDECARDPLLCRGGTCTNTDGSYKCQCPPGHE TAKGTACEDIDECSLSDG CPHGQCVNVIG AFQCSCHAGFQSTPDRQGCVDINECRVQNGGCDVHRINTEGSYRCSCGQGYS MPDGRACADVDECEENPRVCDQGHCTNM PGGHRCLCYDGFMATPDMRTCVDVDECDLNPHICLHGDCENTKGSFVCHCQ GY VRKGATGCΞDVDECEVGGHNCDSHAS CLNIPGSFSCRCLPG VGDGFECHD DECVSQEHRCSPRGDCLNVPGSYRCTCRQGFAGDGFFCEDRDECAENVDLCDNGQ C NAPGGYRCECEMGFDPTEDHRACQDVDECAQENLCAFGSCEN PGMFRCICNGGYELDRGGGNCTDINECADPVNCING VCINTPGSYLCSCPQDFE NPΞGVGCVDTRAGNCFLETHDRGDSGISCSAEIGVGVTRASCCCS GRAWGNPCELCPMANT TEYRT CPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCECPPGYHLSEHTRICEDIDECSTHSG1CGPGT CYNT GNYTCVCPAEYLQVNGGNNCMDMRKSVCFRHYNGTCQNE AFNVTRKMCCCSYNIGQAWNRPCEACPTPISPDYQI LCGNQAPGFLTDIHTGKPLDIDECGEIPAICANGICINQIGSFRCECPAGFNYNSILLACEDVDECGSRESPCQQNADCIN IPGSYRCKCTRGYK SPGGACVGRNECREIPNVCSHGDCMDTEGSYMC CHRGFQASADQTLCMDIDECDRQPCGNGTCKN IIGSYNCLCFPGFVVTHNGDCVDFDECTTLVGQVCRFGHCLNTAGSFHCLCQDGFE TADGKNCVDTNECLSLAGTCLPGT CQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSPGSFQCLCPPGFV SDNGHRCFDTRQSFCFTRFEAGK CSVPKAFNTTKTRCCCΞKRPGEGWGDPCELCPQEDSPPP RPAAFQELCPFGHGAVPGPDDSREDVNECAENPGVCTNGVC VNTDGSFRCECPFGYSLDFTGINCEDTDECSVGHPCGQGTCTNVIGGFECACADGFEPGLMMTCEDIDECS NPLLCAFRC HNTEGSY CTCPAGYTLREDGAMCRDVDECADGQQDCHARGMECKNLIGTFACVCPPGMRPLPGSGEGCTDDNECHAQPD CVNGRCVNTAGSFRCDCDEGFQPSPTLTECHDIRQGPCFAEVLQTMCRSLSSSSEAVTRAECCCGGGRGWGPRCELCPLPG TSAYRK CPHGSGYTAEGRDVDECR LAHLCAHGECINS GSFRCHCQAGYTPDATATTCLDMDECSQVPKPCTFLCKNTK GSFLCSCPRGY EEDGRTCKDLDECTSRQHNCQFLCVNTVGAFTCRCPPGFTQHHQACFDNDECSAQPGPCGAHGHCHNT PGSFRCECHQGFTLVSSGHGCEDVNECDGPHRCQHGCQNQLGGYRCSCPQAFTQHSQ AQCVDENECALSPPTCGSASCRN T GGFRCVCPΞGFDFDQA GGCQEVDECAGRRGPCΞYSCANTPGGF CGCPQGYFRVGQGHCVSGLGFSPGPQDTPDKEEL LSSEACYECKINGLSPRDRPRRSAHRDHQVN ATLDSEALLT GLNLSH GRAERILELRPALEG EGRIRYVIVRGNEQG FFRMHHLRGVSSLQ GRRRPGPGTYRLEVVSHMAGPWGVQQEGQPGPWGQALRLKVQLSVA VGRSLSGPQ SREGGFWNW EGLIPRSDG

The NOV2b amino acid sequence has 1797 of 2750 amino acid residues (65%) identical to, and 2161 of 2750 amino acid residues (78%) similar to, a Homo sapiens 2911 amino acid residue fibrillin 2 precursor protein (ρtnr:SWISSNEW-ACC:P35556) (E = 0.0).

NOV2b is expressed in at least the following tissues: adrenal gland, bone marrow, brain - amygdala, brain - cerebellum, brain - hippocampus, brain - substantia nigra, brain - thalamus, brain -whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma - Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea and uterus. Expression information was derived from the tissue sources ofthe sequences that were included in the derivation ofthe NOV2b sequence.

NOV2c

A disclosed NOV2c nucleic acid of 9993 nucleotides (also referred to as CG88987-01) encoding a novel Fibrillin-like protein is shown in Table 2E. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 520-522 and ending with a TGA codon at nucleotides 9151-9153. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table 2E, and the start and stop codons are in bold letters.

Table 2E. NOV2c nucleotide sequence (SEQ ID NO:9).

CCTCCCATTCCTCTCCTCTCTCTCCCATCTGGACAGCCCCCAGCCTTCTGACACCCTGTTTCCCCCCCGGGACCCTGCTGC CCTTCCCCACACCACCTAACATGCATTTCTGACCTTGCTCCTCCCACTCAAAGCTTTTCAAGGGCTCTTCATTGTCCTTGA CGAATGAGCAAAAGCTGCATGACACTGCTATGCCCACTCCTGCAACGCCTGGCCCCACCTGGCCCCACCTGGCCCTCTCTC CCCTCCCTTCCACTTCCCACTCCCGTCTTGCCTTGGCTGTGCACCCTCAGCCCCAAGACCCTTCTCTAACATCACACCTCC TCCAGGAAGCCTCCTCTGACTGTTCACCGCTTCCCGGTTGGGGCTGGATGCCTCCTTGGGCTCCCAGAGGCCCCGGGGCTG CCCCATTTGGCAACCACCCTGCCCTCCTCGTTGTGTTGGGTCCCCCATGGCTAGGGTGGGGGGCTCATCTAGGGCTGACTC TGCTTCCTCTCCGCAGCCTCCAGGGGACACGCCATGACTCTGGAGGGTCTGTATTTGGCAAGGGGCCCCCTGGCCCGGCTC CTGCTGGCCTGGTCGGCCCTGTTGTGCATGGCAGGTGGCCAAGGCCGCTGGGACGGGGCCTTGGAGGCTGCAGGTCCTGGA CGTGTGCGGAGGCGGGGCAGCCCAGGCATCTTGCAGGGGCCGAATGTGTGCGGCTCCCGGTTCCATGCCTACTGCTGTCCA GGCTGGAGGACATTCCCTGGCAGGAGCCAGTGTGTCGTACCCATCTGTAGGCGCGCCTGCGGTGAAGGCTTCTGCTCCCAG CCCAACCTGTGCACCTGTGCGGATGGGACGCTGGCTCCCAGCTGCGGGGTGAGCCGAGGGTCAGGGTGCAGTGTGAGCTGT ATGAATGGGGGCACCTGCCGGGGGGCGTCCTGTCTGTGTCAGAAGGGCTACACAGGCACCGTGTGTGGGCAGCCCATCTGT GACCGCGGCTGCCACAATGGGGGTCGCTGCATTGGGCCCAACCGCTGCGCCTGTGTGTATGGCTTCATGGGACCTCAATGT GAGAGAGATTACCGGACGGGACCCTGCTTTGGCCAAGTAGGCCCCGAGGGGTGCCAGCATCAGCTGACGGGCCTCGTGTGC ACCAAGGCACTTTGCTGTGCCACTGTGGGCCGTGCCTGGGGCCTTCCATGTGAACTTTGCCCTGCACAGCCACACCCCTGC CGCCGCGGCTTCATCCCCAATATCCACACGGGGGCCTGCCAAGATGTGGATGAGTGCCAGGCTGTGCCAGGCCTGTGCCAG GGAGGCAGCTGCGTCAACATGGTGGGCTCCTTCCATTGCCGCTGTCCAGTTGGACACCGGCTCAGTGACAGCAGCGCCGCA TGTGAAGACTACGATGAATGCAGCACCATTCCTGGAATCTGTGAAGGGGGTGAATGTACAAACACAGTCAGCAGTTACTTT TGCAAATGTCCTCCTGGTTTTTACACCTCTCCAGATGGTACTCTTCATGGACAGTCGCGGGCCGGCGCCTGCTTCTCAGTG CTTTTCGGGGGCCGCTGTGCTGGAGACCTCGCCGGCCACTACACTCGCAGGCAGTGCTGCTGTGACAGGGGCAGGTGCTGG GCAGCTGGCCCGGTCCCTGAGCTGTGTCCTCCTCGGGGCTCCAATGAATTCCAGCAACTGTGCGCCCAGCGGCTGCCGCTG CTACCCGGCCACCCTGGCCTCTTCCCTGGCCTCCTGGGCTTCGGATCCAATGGCATGGGTCCCCCTCTTGGGCCAGCGCGA CTCAACCCCCATGGCTCTGATGCGCGTGGGATCCCCAGCCTGGGCCCTGGCAACTCTAATATTGGCACTGCTACCCTGAAC CAGACCATTGACATCTGCCGACACTTCACCAACCTGTGTCTGAATGGCCGCTGCCTGCCCACGCCTTCCAGCTACCGCTGC GAGTGTAACGTGGGCTACACCCAGGACGTGCGCGGCGAGTGCATTGATGTAGACGAATGCACCAGCAGCCCCTGCCACCAC GGTGACTGCGTCAACATCCCCGGCACCTACCACTGCCGGTGCTACCCGGGCTTCCAGGCCACGCCCACCAGGCAGGCATGC GTGGATGTGGACGAGTGCATTGTCAGTGGTGGCCTTTGTCACCTGGGCCGCTGTGTCAACACAGAGGGCAGCTTCCAGTGT GTCTGCAATGCAGGCTTCGAGCTCAGCCCTGACGGCAAGAACTGTGTGGACCACAACGAGTGTGCCACCAGCACCATGTGC GTCAACGGCGTGTGTCTCAACGAGGATGGCAGCTTCTCCTGCCTCTGCAAACCCGGCTTCCTGCTGGCGCCTGGCGGCCAC TACTGCATGGACATTGACGAGTGCCAGACGCCCGGCATCTGCGTGAACGGCCACTGTACCAACACCGAGGGCTCCTTCCGC TGCCAGTGCCTGGGGGGGCTGGCGGTAGGCACGGATGGCCGCGTGTGCGTGGACACCCACGTGCGCAGCACCTGCTATGGG GCCATCGAGAAGGGCTCCTGTGCCCGCCCCTTCCCTGGCACTGTCACCAAGTCCGAGTGCTGCTGTGCCAATCCGGACCAC GGTTTTGGGGAGCCCTGCCAGCTTTGTCCTGCCAAAGACTCCGCTGAGTTCCAGGCACTGTGCAGCAGTGGGCTTGGCATT ACCACGGATGGTCGAGACATCAACGAGTGTGCTCTGGATCCTGAGGTTTGTGCCAATGGCGTGTGCGAGAACCTTCGGGGC AGCTACCGCTGTGTCTGCAACCTGGGTTATGAGGCAGGTGCCTCAGGCAAGGACTGCACAGACGTGGATGAGTGTGCCCTC AACAGCCTCCTGTGTGACAACGGGTGGTGCCAGAATAGCCCTGGCAGCTACAGCTGCTCCTGCCCCCCCGGCTTCCACTTC TGGCAGGACACGGAGATCTGCAAAGATGTCGACGAATGCCTGTCCAGCCCGTGTGTGAGTGGCGTCTGTCGGAACCTGGCC GGCTCCTACACCTGCAAATGTGGCCCTGGCAGCCGGCTGGACCCCTCTGGTACCTTCTGTCTAGACAGCACCAAGGGCACC TGCTGGCTGAAGATCCAGGAGAGCCGCTGTGAGGTGAACCTTCAGGGAGCCAGCCTGCGGTCTGAGTGCTGTGCCACCCTC GGGGCAGCCTGGGGGAGCCCCTGCGAACGCTGCGAGATCGACCCTGCCTGTGCCCGGGGCTTTGCCCGGATGACGGGTGTC ACCTGCGATGATGTGAACGAGTGTGAGTCCTTCCCGGGAGTCTGTCCCAACGGGCGTTGCGTCAACACTGCTGGGTCTTTC CGCTGTGAGTGTCCAGAGGGCCTGATGCTGGACGCCTCAGGCCGGCTGTGCGTGGATGTGAGATTGGAACCATGTTTCCTG CGATGGGATGAGGATGAGTGTGGGGTCACCCTGCCTGGCAAGTACCGGATGGACGTCTGCTGCTGCTCCATCGGGGCCGTG TGGGGAGTCGAGTGCGAGGCCTGCCCGGATCCCGAGTCTCTGGAGTTCGCCAGCCTGTGCCCGCGGGGGCTGGGCTTCGCC AGCCGGGACTTCCTGTCTGGCCGACCATTCTATAAAGATGTGAATGAATGCAAGGTGTTCCCTGGCCTCTGCACGCACGGT ACCTGCAGAAACACGGTGGGCAGCTTCCACTGCGCCTGTGCGGGGGGCTTCGCCCTGGATGCCCAGGAACGGAACTGCACA GATATCGACGAGTGTCGCATCTCTCCTGACCTCTGCGGCCAGGGCACCTGTGTCAACACGCCGGGCAGCTTTGAGTGCGAG TGTTTTCCCGGCTACGAGAGTGGCTTCATGCTGATGAAGAACTGCATGGACGTGGACGAGTGTGCAAGGGACCCGCTGCTC TGCCGGGGAGGCACTTGCACCAACACGGATGGGAGCTACAAGTGCCAGTGTCCCCCTGGGCATGAGCTGACGGCCAAGGGC ACTGCCTGTGAGGACATCGATGAGTGCTCCCTGAGTGATGGCCTGTGTCCCCATGGCCAGTGTGTCAATGTCATCGGTGCC TTCCAGTGCTCCTGCCATGCCGGCTTCCAGAGCACACCTGACCGCGGAGCTACAAGTGCCAGTTGCCCGACGGAAGGGCAT GTGCAGGTCGTCCTGGGGCCTGGGGAGCAGATGTGTACTGGGTGGTCTATCAGGGCAAAGCTAAGCACAGTCCCCAACCCT GCTCCCCCAGACGTGGACGAGTGTGAAGAGAACCCCCGCGTTTGTGACCAAGGCCACTGCACCAACATGCCAGGGGGTCAC CGCTGCCTGTGCTATGATGGCTTCATGGCCACGCCAGACATGAGGACATGTGTTGATGTGGATGAGTGTGACCTGAACCCT CACATCTGCCTCCATGGGGACTGCGAGAACACGAAGGGTTCCTTTGTCTGCCACTGTCAGCTGGGCTACATGGTCAGGAAG GGGGCCACAGGCTGCTCTGATGTGGATGAATGCGAGGTTGGAGGACACAACTGTGACAGTCACGCCTCCTGTCTCAACATC CCAGGGAGTTTCAGCTGTAGGTGCCTGCCAGGCTGGGTGGGGGATGGCTTCGAATGTCACGACCTGGATGAATGCGTCTCC CAGGAGCACCGGTGCAGCCCAAGAGGTGACTGTCTCAATGTCCCTGGCTCCTACCGCTGCACCTGCCGCCAGGGCTTTGCC GGGGATGGCTTCTTCTGCGAAGACAGGGATGAATGTGCCGAGAACGTGGACCTCTGTGACAACGGGCAGTGCCTCAATGCG CCCGGCGGGTACCGCTGTGAATGTGAGATGGGCTTTGACCCCACCGAGGACCACCGGGCCTGCCAGGATGTGGACGAGTGT GCGCAAGAGAACCTCTGTGCATTTGGGAGCTGTGAGAACCTGCCTGGAATGTTCCGCTGCATCTGCAATGGTGGCTACGAA CTGGACCGAGGGGGTGGCAACTGCACAGACATCAACGAGTGTGCAGACCCAGTAAACTGCATCAACGGCGTGTGCATTAAC ACCCCCGGCAGCTACCTCTGCAGCTGCCCCCAGGATTTTGAGCTGAACCCCAGCGGAGTGGGCTGCGTGGACACTCGGGCC GGGAACTGTTTCCTGGAGACGCATGACCGAGGGGACAGTGGCATTTCCTGCAGTGCCGAGATCGGAGTTGGTGTCACCCGA GCTTCCTGCTGTTGCTCCCTGGGCCGGGCTTGGGGCAATCCCTGTGAGCTGTGCCCTATGGCCAACACCACTGAGTACAGA ACCCTGTGCCCGGGTGGTGAGGGCTTCCAGCCTAACCGCATCACTGTCATTCTGGAAGACATCGACGAGTGCCAAGAGCTG CCAGGGCTGTGTCAGGGGGGTGACTGCGTCAACACGTTTGGCAGTTTCCAGTGTGAGTGCCCACCTGGCTACCACCTCAGT GAGCACACCCGCATCTGTGAGGATATTGACGAATGCTCCACACACTCCGGCATCTGTGGCCCTGGCACCTGCTACAACACC CTGGGGAACTACACCTGTGTCTGCCCTGCAGAGTACCTCCAAGTCAATGGTGGCAACAACTGCATGGATATGAGGAAGAGT GTCTGCTTCCGGCACT_ATAACGGCACATGTCA_AAATG_AGCTGGCCTTCAACGTGACCCGGAAAATGTGTTGCTGCTCCTAC A_AC_ATTGGCCAGGCCTGGA_ATAGACCCTGTGAGGCCTGCCCCACTCCCATCAGTCCTGACTACCAGATCCTGTGTGGAAAT C_AGGCCCCGGG_ATTCCTCACTG_AC_ATCC_ACACGGGGA_AGCCCCTTGACATTGATGAGTGTGGGGAGATCCCCGCCATCTGT GCC_AATGGCATCTGC_AT_AAACC_AGATCGGGAGTTTCCGCTGCGAGTGCCCCGCAGGCTTCAACTACAACAGCATCCTGCTG GCTTGTG_AAG_ATGTCG_ATG_AGTGTGGCAGCAGGGAGAGTCCCTGCCAGCAGAATGCTGACTGCATCAACATCCCCGGTAGC T_ACCGCTGCAAGTGC_ACCCG_AGGGTACAAACTGTCGCCAGGCGGGGCTTGTGTGGGACGGAATGAGTGTCGGGAGATCCCG _AATGTCTGTAGCC_ATGGTG_ACTGCATGGACACAG_AAGGCAGCTACATGTGTCTGTGTCACCGTGGATTCCAGGCCTCTGCA GACCAG_ACCCTGTGCATGG_AC_ATTGACGAGTGTG_ACCGGCAGCCTTGTGGAAATGGGACCTGCAAGAACATCATTGGCTCC T_ACA_ACTGCCTCTGCTTCCCTGGCTTTGTGGTGACACACAATGGGGATTGTGTGGATTTTGATGAGTGTACTACCCTGGTG GGGCAGGTGTGCCGATTTGGCC_ATTGCCTC_AAC_AC_AGCTGGTTCCTTCCACTGCCTCTGCCAGGATGGCTTTGAGCTCACA GCTG_ATGGG_AAGA_ACTGTGTGG_ACACCAATG_AGTGCCTCAGCCTTGCAGGAACCTGCCTACCCGGCACTTGCCAGAACCTC G_AGGGCTCCTTCCGCTGCATCTGTCCCCCTGGCTTCCAGGTGCAGAGTGACCACTGCATTGATATCGACGAGTGCTCAGAG G_AGCCC_AACCTCTGCCTCTTTGGCACCTGT_ACCAACAGCCCTGGGAGCTTCCAGTGCCTCTGCCCACCTGGCTTTGTCCTC TCTG_AC_AATGGGC_ACCGTTGCTTTGACACACGGCAGAGTTTCTGCTTCACCCGTTTTGAGGCTGGGAAGTGCTCGGTGCCC _AAAGCTTTCA_AC_ACC_ACC_AAG_ACCCGCTGCTGCTGC_AGT_AAGAGGCCTGGGGAGGGCTGGGGAGACCCCTGCGAACTGTGT CCCC_AGGA_AGGCAGCGCTGCCTTTCAGGAGCTCTGCCCCTTTGGCCACGGGGCAGTCCCAGGCCCGGATGACTCCCGAGAA G_ACGTG_AATG_AGTGTGCAGAG_AACCCTGGCGTCTGCACTAACGGCGTCTGTGTCAACACCGATGGATCCTTCCGCTGTGAG TGTCCCTTTGGCT_ACAGCCTGGACTTCACTGGCATCAACTGTGTGGACACAGACGAGTGCTCTGTCGGCCACCCCTGTGGG C_AAGGG_AC_ATGC_ACC_AATGTCATCGGAGGCTTCGAATGTGCCTGTGCTGACGGCTTTGAGCCTGGCCTCATGATGACCTGC GA_GGAC_ATCG_ACG_AATGCTCCCTGAACCCGCTGCTCTGTGCCTTCCGCTGCCACAATACCGAGGGCTCCTACCTGTGCACC TGTCCAGCCGGCT_ACACCCTGCGGGAGGATGGGGCCATGTGTCGAGATGTGGACGAGTGTGCAGATGGTCAGCAGGACTGC C_ACGCCCGGGGC_ATGGAGTGCAAGAACCTCATCGGTACCTTCGCGTGCGTCTGTCCCCCAGGCATGCGGCCCCTGCCTGGC TCTGGGG_AGGGCTGCACAGATGACAATGAATGCCACGCTCAGCCTGACCTCTGTGTCAACGGCCGCTGTGTCAACACCGCG GGC_AGCTTCCGGTGCG_ACTGTG_ATGAGGGATTCCAGCCCAGCCCCACCCTTACCGAGTGCCACGACATCCGGCAGGGGCCC TGCTTTGCCG_AGGTGCTG_CAGACCATGTGCCGGTCTCTGTCCAGCAGCAGTGAGGCTGTCACCAGGGCCGAGTGCTGCTGT GGGGGTGGCCG_GGGCT_GGGGGCCCCGCTGCGAGCTCTGTCCCCTGCCCGGCACCTCTGCCTACAGGAAGCTGTGCCCCCAT GGCTCAGGCTACACTGCTGAGGGCCGAGATGTAGATGAATGCCGTATGCTTGCTCACCTGTGTGCTCATGGGGAGTGCATC _AACAGCCTTGGCTCCTTCCGCTGCCACTGTCAGGCCGGGTACACACCGGATGCTACTGCTACTACCTGCCTGGATATGGAT GAGTGCAGCCAGGTCCCCAAGCCATGTACCTTCCTCTGCAAAAACACGAAGGGCAGTTTCCTGTGCAGCTGTCCCCGAGGC TACCTGCTGGAGGAGGATGGCAGGACCTGCAAAGACCTGGACGAATGCACCTCCCGGCAGCACAACTGTCAGTTCCTCTGT GTCAACACTGTGGGCGCCTTCACCTGCCGCTGTCCACCCGGCTTCACCCAGCACCACCAGGCCTGCTTCGACAATGATGAG TGCTCAGCCCAGCCTGGCCCATGTGGTGCCCACGGGCACTGCCACAACACCCCGGGCAGCTTCCGCTGTGAATGCCACCAA GGCTTCACCCTGGTCAGCTCAGGCCATGGCTGTGAAGATGTGAATGAATGTGATGGGCCCCACCGCTGCCAGCATGGCTGT CAGAACCAGCTAGGGGGCTACCGCTGCAGCTGCCCCCAGGCTTTCACCCAGCACTCCCAGTGGGCCCAGTGTGTGGATGAG AATGAGTGTGCCCTGTCGCCCCCCACCTGCGGGAGCGCCTCCTGTCGCAACACTCTTGGTGGCTTCCGCTGCGTCTGCCCC TCCGGCTTTGACTTTGATCAGGCCCTCGGGGGCTGCCAGGAGGTGGATGAGTGCGCCGGACGGCGTGGCCCCTGTAGCTAC AGCTGTGCCAACACGCCTGGTGGCTTCCTGTGCGGCTGTCCTCAAGGCTACTTCCGGGTTGGGCAAGGGCACTGTGTCTCC GGCCTGGGCTTCAGCCCCGGACCCCAGGACACCCCGGACAAAGAGGAGCTGCTCTCGTCTGAAGCCTGCTACGAATGCAAG ATCAATGGCCTCTCCCCTCGGGACCGGCCACGACGCAGTGCCCACAGGGACCACCAGGTGAACCTGGCCACCCTTGACTCC GAGGCCCTGCTGACCTTGGGCCTGAACCTCTCACACCTGGGCCGGGCCGAGCGCATCCTGGAGCTCCGGCCGGCCCTGGAG GGTCTAGAGGGCCGGATCCGCTACGTCATCGTCCGCGGAAACGAGCAAGGTTTCTTTCGCATGCATCACCTCCGTGGCGTC AGCTCCCTGCAGCTGGGGCGGAGGCGGCCGGGGCCTGGAACCTACCGGCTGGAGGTGGTGAGCCACATGGCAGGACCCTGG GGTGTCCAGCAAGAGGGGCAGCCAGGGCCATGGGGCCAGGCCTTGAGGCTGAAGGTGCAACTGTCAGTTGCTTTAGTTGGG AGGAGCCTCAGTGGGCCCCAGCTGTCCAGAGAAGGGGGATTCTGGAACTGGGAAGGACTGATCCCCAGAAGCGATGGCTGA CCAGATTGAACCCCGAAACTCAGGAAGAGTGAAATGCTACACGACAACCTCAGGCAAGCCCGGCCTCTGCCTGGGCCTCTG TGCCAGCCCCGGGGGCCCCCCAGTTACTCAGTCTTTCCTGGAGACAGCAAGAAGCTGCAATGTGCAATCCCCCTGCCCCCA CAGCCAAGGTCAGGAAGAGGCCCTGTGGTCACCGTGTCTGGCCAATCTCAGGCTTTCACTTCTGTACTGCACTGTGGCTTG CCCTGGCGGGGGGCAGGGGGTTGGCAGGACATGGCAATGGGCAACTGGGGTGGGCACAGGGCTTATTCCTCGGAGTAGAAG GGTGTACAGGGGGCCCAGACTCCACAGTGACTTGCCACΆTTTGCCCCCCATTTGGΆGAATGCTTTTATATCAAAAGTGGAG ACGATAATAAAGTTATTTTGGGTTAAGTCTGCCTGCCCTTTGGCAAGTTCTTGAAGTAAGTAGATGCTGCCCTCGGACTGG GCGAGGCAGATCTTGTGCCTGGGGAAGCAGAAGGCCTTATGGGCTCCCCAGAATGGTAATAATGGCTCACGCTTCCTGACC ACGTACTACATACCAGΆCACCATTCGATTTTTTTTTTATTTTTTCTGAGACAGGGTCTTGCTCTGTTGCCTAGGTTGGAGT GCAGTGGCGCGATCGTGGCTCCCTGTGGTCGCCACTTTCCGGGCTCGAGCAATCCTCCCACCTCAGCCTCTCCCAAGTAGC TGGGACCAAAGGTGCACGCCACCACACCCAGCTAATTTTTTTAAATTTCTTTTTCTTTCTCTTTCTTTTTGAGACCAGCCC GGCCAACATGACGAAACCCCATCTCTAATA

The disclosed NOV2c nucleic acid sequence, localized to chromsome 19, has 3194 of 4382 bases (72%) identical to a Mus musculus fibrillin 2 (fbn2) mRNA (gb:GENBANK- ID:MUSFBN2|acc:L39790.1) (E = 0.0).

A NOV2c polypeptide (SEQ ID NO: 10) encoded by SEQ ID NO:9 has 2877 amino acid residues and is presented using the one-letter code in Table 2F. Signal P, Psort and/or Hydropathy results predict that NOV2c contains a signal peptide and is likely to be localized to the nucleus with a certainty of 0.6000. Although PSORT suggests that the Fibrillin-like protein may be localized in the nucleus, the NOV2c protein is similar to the Fibrillin family, some members of which are released extracellularly. Therefore it is likely that NOV2c protein shows a similar localization. The most likely cleavage site for a NOV2c peptide is between amino acids 29 and 30, at: AGG-QG.

Table 2F. Encoded NOV2c protein sequence (SEQ ID NO:10).

MTLEGLYLARGP ARL LAWSALLCMAGGQGRWDGALEAAGPGRVRRRGSPGILQGPNVCGSRFHAYCCPGWRTFPGRSQC VVPICRRACGEGFCSQPNLCTCADGT APSCGVSRGSGCSVSCMNGGTCRGASCLCQKGYTGTVCGQPICDRGCHNGGRCI GPNRCACVYGFMGPQCERDYRTGPCFGQVGPEGCQHQ TGLVCTKALCCATVGRAWGLPCE CPAQPHPCRRGFIPNIHTG ACQDVDECQAVPGLCQGGSCVNMVGΞFHCRCPVGHR SDSSAACEDYDECSTIPGICEGGECTNTVSSYFCKCPPGFYTSP DGTLHGQSRAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSNEFQQLCAQR PLLPGHPGLFPGL LGFGSNGMGPP GPARLNPHGSDARGIPS GPGNSNIGTATLNQTIDICRHFTNLCLNGRC PTPSSYRCECNVGYTQDVR GECIDVDECTSSPCHHGDCVNIPGTYHCRCYPGFQATPTRQACVDVDECIVΞGGLCH GRCVNTEGSFQCVCNAGFELSPD GKNCVDHNECATST CVNGVCLNEDGSFSC CKPGFLLAPGGHYCMDIDECQTPGICVNGHCTNTEGSFRCQCLGGLAVGT DGRVCVDTHVRSTCYGAIEKGSCARPFPGTVTKSECCCANPDHGFGEPCQLCPAKDSAEFQALCSSGLGITTDGRDINECA LDPEVCANGVCENLRGSYRCVCNLGYEAGASGKDCTDVDECA NSLLCDNGWCQNSPGSYSCSCPPGFHFWQDTEICKDVD ECLSSPCVSGVCRNLAGSYTCKCGPGSRLDPSGTFC DSTKGTCWLKIQESRCEVNLQGASLRSECCAT GAAWGΞPCERC EIDPACARGFARMTGVTCDDVNECESFPGVCPNGRCVNTAGSFRCECPEGLMLDASGRLCVDVR EPCFLRWDEDECGVTL PGKYRMDVCCCSIGAVWGVECEACPDPES EFASLCPRGLGFASRDFLSGRPFYKDVNECKVFPGLCTHGTCRNTVGSFHC ACAGGFALDAQERNCTDIDECRISPD CGQGTCVNTPGΞFECECFPGYESGF LMKNCMDVDECARDP LCRGGTCTNTDG SYKCQCPPGHELTAKGTACEDIDECS SDG CPHGQCVNVIGAFQCSCHAGFQSTPDRGATSASCPTEGHVQVVLGPGEQM CTGWSIRAKLSTVPNPAPPDVDECEENPRVCDQGHCTNMPGGHRCLCYDGFMATPDMRTCVDVDECDLNPHICLHGDCENT KGSFVCHCQ GY VRKGATGCΞDVDECEVGGHNCDΞHASCLNIPGSFSCRCLPG VGDGFECHDLDECVSQEHRCSPRGDC LNVPGSYRCTCRQGFAGDGFFCEDRDECAENVD CDNGQCLNAPGGYRCECEMGFDPTEDHRACQDVDECAQENLCAFGSC ENLPGMFRCICNGGYE DRGGGNCTDINECADPVNCINGVCINTPGSYLCSCPQDFELNPSGVGCVDTRAGNCFLETHDRG DSGISCSAEIGVGVTRASCCCSLGRA GNPCELCPMANTTEYRTLCPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVN TFGSFQCECPPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTLGNYTCVCPAEYLQVNGGNNC DMRKSVCFRHYNGTCQ NELAFNVTRKMCCCSYNIGQAWNRPCEACPTPISPDYQILCGNQAPGFLTDIHTGKPLDIDECGEIPAICANGICINQIGS FRCECPAGFNYNSILLACEDVDECGSRESPCQQNADCINIPGSYRCKCTRGYK SPGGACVGRNECREIPNVCSHGDC DT EGSYMCLCHRGFQASADQTLCMDIDECDRQPCGNGTCKNIIGSYNCLCFPGFVVTHNGDCVDFDECTTLVGQVCRFGHCLN TAGSFHCLCQDGFELTADGKNCVDTNECLSLAGTCLPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCT NSPGSFQCLCPPGFV SDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRCCCSKRPGEGWGDPCELCPQEGSAAFQEL CPFGHGAVPGPDDSREDVNECAENPGVCTNGVCVNTDGSFRCECPFGYSLDFTGINCVDTDECSVGHPCGQGTCTNVIGGF ECACADGFEPGL MTCEDIDECSLNPLLCAFRCHNTEGSYLCTCPAGYT REDGAMCRDVDECADGQQDCHARG ECKNLI GTFACVCPPGMRPLPGSGEGCTDDNECHAQPDLCVNGRCVNTAGSFRCDCDEGFQPSPTLTECHDIRQGPCFAEVLQTMCR SLSSSSEAVTRAECCCGGGRGWGPRCELCP PGTSAYRKLCPHGSGYTAEGRDVDECRMLAHLCAHGECINSLGSFRCHCQ AGYTPDATATTCLDMDECΞQVPKPCTFLCKNTKGSF CSCPRGYLLEEDGRTCKDLDECTSRQHNCQFLCVNTVGAFTCRC PPGFTQHHQACFDNDECSAQPGPCGAHGHCHNTPGSFRCECHQGFTLVSSGHGCEDVNECDGPHRCQHGCQNQLGGYRCSC PQAFTQHSQ AQCVDENECALSPPTCGSASCRNTLGGFRCVCPSGFDFDQA GGCQEVDECAGRRGPCSYSCANTPGGF C GCPQGYFRVGQGHCVSG GFSPGPQDTPDKEE SSEACYECKINGLSPRDRPRRSAHRDHQVNLATLDSEA LTLGLNLS H GRAERILELRPALEGLEGRIRYVIVRGNEQGFFR HHLRGVSSLQ GRRRPGPGTYRLEVVSH AGP GVQQEGQPGPW GQALRLKVQLSVA VGRSLSGPQLSREGGF N EG IPRSDG The NOV2c amino acid sequence has 1897 of 2810 amino acid residues (67%) identical to, and 2258 of 2810 amino acid residues (80%) similar to, a Mus musculus 2907 amino acid residue fibrillin 2 precursor protein (ptnr:SWISSPROT-ACC:Q61555) (E = 0.0).

NOV2c is expressed in at least the following tissues: adrenal gland, bone marrow, brain - amygdala, brain - cerebellum, brain - hippocampus, brain - substantia nigra, brain - thalamus, brain -whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma - Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea and uterus. Expression information was derived from the tissue sources ofthe sequences that were included in the derivation ofthe NOV2c sequence. NOV2d

A disclosed NOV2d nucleic acid of 9418 nucleotides (also referred to as CG88987-02) encoding a novel Fibrillin-like protein is shown in Table 2G. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 41-43 and ending with a TGA codon at nucleotides 8576-8578. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table 2G, and the start and stop codons are in bold letters.

Table 2G. NOV2d nucleotide sequence (SEQ ID NO:ll).

CTGACTCTGCTTCCTCTCCGCAGCCTCCAGGGGACACGCCATGACTCTGGAGGGTCTGTATTTGGCAAGGGGCCCCCTGGC CCGGCTCCTGCTGGCCTGGTCGGCCCTGTTGTGCATGGCAGGTGGCCAAGGCCGCTGGGACGGGGCCTTGGAGGCTGCAGG TCCTGGACGTGTGCGGAGGCGGGGCAGCCCAGGCATCTTGCAGGGGCCGAATGTGTGCGGCTCCCGGTTCCATGCCTACTG CTGTCCAGGCTGGAGGACATTCCCTGGCAGGAGCCAGTGTGTCGTACCCATCTGTAGGCGCGCCTGCGGTGAAGGCTTCTG CTCCCAGCCCAACCTGTGCACCTGTGCGGATGGGACGCTGGCTCCCAGCTGCGGGGTGAGCCGAGCCATCTGTGACCGCGG CTGCCACAATGGGGGTCGCTGCATTGGGCCCAACCGCTGCGCCTGTGTGTATGGCTTCATGGGACCTCAATGTGAGAGAGA TTACCGGACGGGACCCTGCTTTGGCCAAGTAGGCCCCGAGGGGTGCCAGCATCAGCTGACGGGCCTCGTGTGCACCAAGGC ACTTTGCTGTGCCACTGTGGGCCGTGCCTGGGGCCTTCCATGTGAACTTTGCCCTGCACAGCCACACCCCTGCCGCCGCGG CTTCATCCCCAATATCCACACGGGGGCCTGCCAAGATGTGGATGAGTGCCAGGCTGTGCCAGGCCTGTGCCAGGGAGGCAG CTGCGTCAACATGGTGGGCTCCTTCCATTGCCGCTGTCCAGTTGGACACCGGCTCAGTGACAGCAGCGCCGCATGTGAAGA CTACGATGAATGCAGCACCATTCCTGGAATCTGTGAAGGGGGTGAATGTACAAACACAGTCAGCAGTTACTTTTGCAAATG TCCTCCTGGTTTTTACACCTCTCCAGATGGTACTCTTCATGGACAGTCGCGGGCCGGCGCCTGCTTCTCAGTGCTTTTCGG GGGCCGCTGTGCTGGAGACCTCGCCGGCCACTACACTCGCAGGCAGTGCTGCTGTGACAGGGGCAGGTGCTGGGCAGCTGG CCCGGTCCCTGAGCTGTGTCCTCCTCGGGGCTCCAATGAATTCCAGCAACTGTGCGCCCAGCGGCTGCCGCTGCTACCCGG CCACCCTGGCCTCTTCCCTGGCCTCCTGGGCTTCGGATCCAATGGCATGGGTCCCCCTCTTGGGCCAGCGCGACTCAACCC CCATGGCTCTGATGCGCGTGGGATCCCCAGCCTGGGCCCTGGCAACTCTAATATTGGCACTGCTACCCTGAACCAGACCAT TGACATCTGCCGACACTTCACCAACCTGTGTCTGAATGGCCGCTGCCTGCCCACGCCTTCCAGCTACCGCTGCGAGTGTAA CGTGGGCTACACCCAGGACGTGCGCGGCGAGTGCATTGATGTAGACGAATGCACCAGCAGCCCCTGCCACCACGGTGACTG CGTCAACATCCCCGGCACCTACCACTGCCGGTGCTACCCGGGCTTCCAGGCCACGCCCACCAGGCAGGCATGCGTGGATGT GGACGAGTGCATTGTCAGTGGTGGCCTTTGTCACCTGGGCCGCTGTGTCAACACAGAGGGCAGCTTCCAGTGTGTCTGCAA TGCAGGCTTCGAGCTCAGCCCTGACGGCAAGAACTGTGTGGACCACAACGAGTGTGCCACCAGCACCATGTGCGTCAACGG CGTGTGTCTCAACGAGGATGGCAGCTTCTCCTGCCTCTGCAAACCCGGCTTCCTGCTGGCGCCTGGCGGCCACTACTGCAT GGACATTGACGAGTGCCAGACGCCCGGCATCTGCGTGAACGGCCACTGTACCAACACCGAGGGCTCCTTCCGCTGCCAGTG CCTGGGGGGGCTGGCGGTAGGCACGGATGGCCGCGTGTGCGTGGACACCCACGTGCGCAGCACCTGCTATGGGGCCATCGA GAAGGGCTCCTGTGCCCGCCCCTTCCCTGGCACTGTCACCAAGTCCGAGTGCTGCTGTGCCAATCCGGACCACGGTTTTGG GGAGCCCTGCCAGCTTTGTCCTGCCAAAGACTCCGCTGAGTTCCAGGCACTGTGCAGCAGTGGGCTTGGCATTACCACGGA TGGTCGAGACATCAACGAGTGTGCTCTGGATCCTGAGGTTTGTGCCAATGGCGTGTGCGAGAACCTTCGGGGCAGCTACCG CTGTGTCTGCAACCTGGGTTATGAGGCAGGTGCCTCAGGCAAGGACTGCACAGACGTGGATGAGTGTGCCCTCAACAGCCT CCTGTGTGACAACGGGTGGTGCCAGAATAGCCCTGGCAGCTACAGCTGCTCCTGCCCCCCCGGCTTCCACTTCTGGCAGGA CACGGAGATCTGCAAAGATGTCGACGAATGCCTGTCCAGCCCGTGTGTGAGTGGCGTCTGTCGGAACCTGGCCGGCTCCTA CACCTGCAAATGTGGCCCTGGCAGCCGGCTGGACCCCTCTGGTACCTTCTGTCTAGACAGCACCAAGGGCACCTGCTGGCT GAAGATCCAGGAGAGCCGCTGTGAGGTGAACCTTCAGGGAGCCAGCCTGCGGTCTGAGTGCTGTGCCACCCTCGGGGCAGC CTGGGGGAGCCCCTGCGAACGCTGCGAGATCGACCCTGCCTGTGCCCGGGGCTTTGCCCGGATGACGGGTGTCACCTGCGA TGATGTGAACGAGTGTGAGTCCTTCCCGGGAGTCTGTCCCAACGGGCGTTGCGTCAACACTGCTGGGTCTTTCCGCTGTGA GTGTCCAGAGGGCCTGATGCTGGACGCCTCAGGCCGGCTGTGCGTGGATGTGAGATTGGAACCATGTTTCCTGCGATGGGA TGAGGATGAGTGTGGGGTCACCCTGCCTGGCAAGTACCGGATGGACGTCTGCTGCTGCTCCATCGGGGCCGTGTGGGGAGT CGAGTGCGAGGCCTGCCCGGATCCCGAGTCTCTGGAGTTCGCCAGCCTGTGCCCGCGGGGGCTGGGCTTCGCCAGCCGGGA CTTCCTGTCTGGCCGACCATTCTATAAAGATGTGAATGAATGCAAGGTGTTCCCTGGCCTCTGCACGCACGGTACCTGCAG AAACACGGTGGGCAGCTTCCACTGCGCCTGTGCGGGGGGCTTCGCCCTGGATGCCCAGGAACGGAACTGCACAGATATCGA CGAGTGTCGCATCTCTCCTGACCTCTGCGGCCAGGGCACCTGTGTCAACACGCCGGGCAGCTTTGAGTGCGAGTGTTTTCC CGGCTACGAGAGTGGCTTCATGCTGATGAAGAACTGCATGGACGTGGACGAGTGTGCAAGGGACCCGCTGCTCTGCCGGGG AGGCACTTGCACCAACACGGATGGGAGCTACAAGTGCCAGTGTCCCCCTGGGCATGAGCTGACGGCCAAGGGCACTGCCTG TGAGGACATCGATGAGTGCTCCCTGAGTGATGGCCTGTGTCCCCATGGCCAGTGTGTCAATGTCATCGGTGCCTTCCAGTG CTCCTGCCATGCCGGCTTCCAGAGCACACCTGACCGCGGAGCTACAAGTGCCAGTTGCCCGACGGAAGGGCATGTGCAGGT CGTCCTGGGGCCTGGGGAGCAGATGTGTACTGGGTGGTCTATCAGGGCAAAGCTAAGCACAGTCCCCAACCCTGCTCCCCC AGACGTGGACGAGTGTGAAGAGAACCCCCGCGTTTGTGACCAAGGCCACTGCACCAACATGCCAGGGGGTCACCGCTGCCT GTGCTATGATGGCTTCATGGCCACGCCAGACATGAGGACATGTGTTGATGTGGATGAGTGTGACCTGAACCCTCACATCTG CCTCCATGGGGACTGCGAGAACACGAAGGGTTCCTTTGTCTGCCACTGTCAGCTGGGCTACATGGTCAGGAAGGGGGCCAC AGGCTGCTCTGATGTGGATGAATGCGAGGTTGGAGGACACAACTGTGACAGTCACGCCTCCTGTCTCAACATCCCAGGGAG TTTCAGCTGTAGGTGCCTGCCAGGCTGGGTGGGGGATGGCTTCGAATGTCACGACCTGGATGAATGCGTCTCCCAGGAGCA CCGGTGCAGCCCAAGAGGTGACTGTCTCAATGTCCCTGGCTCCTACCGCTGCACCTGCCGCCAGGGCTTTGCCGGGGATGG CTTCTTCTGCGAAGACAGGGATGAATGTGCCGAGAACGTGGACCTCTGTGACAACGGGCAGTGCCTCAATGCGCCCGGCGG GTACCGCTGTGAATGTGAGATGGGCTTTGACCCCACCGAGGACCACCGGGCCTGCCAGGATGTGGACGAGTGTGCGCAAGA GAACCTCTGTGCATTTGGGAGCTGTGAGAACCTGCCTGGAATGTTCCGCTGCATCTGCAATGGTGGCTACGAACTGGACCG AGGGGGTGGCAACTGCACAGACATCAACGAGTGTGCAGACCCAGTAAACTGCATCAACGGCGTGTGCATTAACACCCCCGG CAGCTACCTCTGCAGCTGCCCCCAGGATTTTGAGCTGAACCCCAGCGGAGTGGGCTGCGTGGACACTCGGGCCGGGAACTG TTTCCTGGAGACGCATGACCGAGGGGACAGTGGCATTTCCTGCAGTGCCGAGATCGGAGTTGGTGTCACCCGAGCTTCCTG CTGTTGCTCCCTGGGCCGGGCTTGGGGCAATCCCTGTGAGCTGTGCCCTATGGCCAACACCACTGAGTACAGAACCCTGTG CCCGGGTGGTGAGGGCTTCCAGCCTAACCGCATCACTGTCATTCTGGAAGACATCGACGAGTGCCAAGAGCTGCCAGGGCT GTGTCAGGGGGGTGACTGCGTCAACACGTTTGGCAGTTTCCAGTGTGAGTGCCCACCTGGCTACCACCTCAGTGAGCACAC CCGCATCTGTGAGGATATTGACGAATGCTCCACACACTCCGGCATCTGTGGCCCTGGCACCTGCTACAACACCCTGGGGAA CTACACCTGTGTCTGCCCTGCAGAGTACCTCCAAGTCAATGGTGGCAACAACTGCATGGATATGAGGAAGAGTGTCTGCTT CCGGCACTATAACGGCACATGTCAAAATGAGCTGGCCTTCAACGTGACCCGGAAAATGTGTTGCTGCTCCTACAACATTGG CCAGGCCTGGAATAGACCCTGTGAGGCCTGCCCCACTCCCATCAGTCCTGACTACCAGATCCTGTGTGGAAATCAGGCCCC GGGATTCCTCACTGACATCCACACGGGGAAGCCCCTTGACATTGATGAGTGTGGGGAGATCCCCGCCATCTGTGCCAATGG CATCTGCATAAACCAGATCGGGAGTTTCCGCTGCGAGTGCCCCGCAGGCTTCAACTACAACAGCATCCTGCTGGCTTGTGA AGATGTCGATGAGTGTGGCAGCAGGGAGAGTCCCTGCCAGCAGAATGCTGACTGCATCAACATCCCCGGTAGCTACCGCTG CAAGTGCACCCGAGGGTACAAACTGTCGCCAGGCGGGGCTTGTGTGGGACGGAATGAGTGTCGGGAGATCCCGAATGTCTG TAGCCATGGTGACTGCATGGACACAGAAGGCAGCTACATGTGTCTGTGTCACCGTGGATTCCAGGCCTCTGCAGACCAGAC CCTGTGCATGGACATTGACGAGTGTGACCGGCAGCCTTGTGGAAATGGGACCTGCAAGAACATCATTGGCTCCTACAACTG CCTCTGCTTCCCTGGCTTTGTGGTGACACACAATGGGGATTGTGTGGATTTTGATGAGTGTACTACCCTGGTGGGGCAGGT GTGCCGATTTGGCCATTGCCTCAACACAGCTGGTTCCTTCCACTGCCTCTGCCAGGATGGCTTTGAGCTCACAGCTGATGG GAAGAACTGTGTGGACACCAATGAGTGCCTCAGCCTTGCAGGAACCTGCCTACCCGGCACTTGCCAGAACCTCGAGGGCTC CTTCCGCTGCATCTGTCCCCCTGGCTTCCAGGTGCAGAGTGACCACTGCATTGATATCGACGAGTGCTCAGAGGAGCCCAA CCTCTGCCTCTTTGGCACCTGTACCAACAGCCCTGGGAGCTTCCAGTGCCTCTGCCCACCTGGCTTTGTCCTCTCTGACAA TGGGCACCGTTGCTTTGACACACGGCAGAGTTTCTGCTTCACCCGTTTTGAGGCTGGGAAGTGCTCGGTGCCCAAAGCTTT CAACACCACCAAGACCCGCTGCTGCTGCAGTAAGAGGCCTGGGGAGGGCTGGGGAGACCCCTGCGAACTGTGTCCCCAGGA AGGCAGCGCTGCCTTTCAGGAGCTCTGCCCCTTTGGCCACGGGGCAGTCCCAGGCCCGGATGACTCCCGAGAAGACGTGAA TGAGTGTGCAGAGAACCCTGGCGTCTGCACTAACGGCGTCTGTGTCAACACCGATGGATCCTTCCGCTGTGAGTGTCCCTT TGGCTACAGCCTGGACTTCACTGGCATCAACTGTGTGGACACAGACGAGTGCTCTGTCGGCCACCCCTGTGGGCAAGGGAC ATGCACCAATGTCATCGGAGGCTTCGAATGTGCCTGTGCTGACGGCTTTGAGCCTGGCCTCATGATGACCTGCGAGGACAT CGACGAATGCTCCCTGAACCCGCTGCTCTGTGCCTTCCGCTGCCACAATACCGAGGGCTCCTACCTGTGCACCTGTCCAGC CGGCTACACCCTGCGGGAGGATGGGGCCATGTGTCGAGATGTGGACGAGTGTGCAGATGGTCAGCAGGACTGCCACGCCCG GGGCATGGAGTGCAAGAACCTCATCGGTACCTTCGCGTGCGTCTGTCCCCCAGGCATGCGGCCCCTGCCTGGCTCTGGGGA GGGCTGCACAGATGACAATGAATGCCACGCTCAGCCTGACCTCTGTGTCAACGGCCGCTGTGTCAACACCGCGGGCAGCTT CCGGTGCGACTGTGATGAGGGATTCCAGCCCAGCCCCACCCTTACCGAGTGCCACGACATCCGGCAGGGGCCCTGCTTTGC CGAGGTGCTGCAGACCATGTGCCGGTCTCTGTCCAGCAGCAGTGAGGCTGTCACCAGGGCCGAGTGCTGCTGTGGGGGTGG CCGGGGCTGGGGGCCCCGCTGCGAGCTCTGTCCCCTGCCCGGCACCTCTGCCTACAGGAAGCTGTGCCCCCATGGCTCAGG CTACACTGCTGAGGGCCGAGATGTAGATGAATGCCGTATGCTTGCTCACCTGTGTGCTCATGGGGAGTGCATCAACAGCCT TGGCTCCTTCCGCTGCCACTGTCAGGCCGGGTACACACCGGATGCTACTGCTACTACCTGCCTGGATATGGATGAGTGCAG CCAGGTCCCCAAGCCATGTACCTTCCTCTGCAAAAACACGAAGGGCAGTTTCCTGTGCAGCTGTCCCCGAGGCTACCTGCT GGAGGAGGATGGCAGGACCTGCAAAGACCTGGACGAATGCACCTCCCGGCAGCACAACTGTCAGTTCCTCTGTGTCAACAC TGTGGGCGCCTTCACCTGCCGCTGTCCACCCGGCTTCACCCAGCACCACCAGGCCTGCTTCGACAATGATGAGTGCTCAGC CCAGCCTGGCCCATGTGGTGCCCACGGGCACTGCCACAACACCCCGGGCAGCTTCCGCTGTGAATGCCACCAAGGCTTCAC CCTGGTCAGCTCAGGCCATGGCTGTGAAGATGTGAATGAATGTGATGGGCCCCACCGCTGCCAGCATGGCTGTCAGAACCA GCTAGGGGGCTACCGCTGCAGCTGCCCCCAGGCTTTCACCCAGCACTCCCAGTGGGCCCAGTGTGTGGATGAGAATGAGTG TGCCCTGTCGCCCCCCACCTGCGGGAGCGCCTCCTGTCGCAACACTCTTGGTGGCTTCCGCTGCGTCTGCCCCTCCGGCTT TGACTTTGATCAGGCCCTCGGGGGCTGCCAGGAGGTGGATGAGTGCGCCGGACGGCGTGGCCCCTGTAGCTACAGCTGTGC CAACACGCCTGGTGGCTTCCTGTGCGGCTGTCCTCAAGGCTACTTCCGGGTTGGGCAAGGGCACTGTGTCTCCGGCCTGGG CTTCAGCCCCGGACCCCAGGACACCCCGGACAAAGAGGAGCTGCTCTCGTCTGAAGCCTGCTACGAATGCAAGATCAATGG CCTCTCCCCTCGGGACCGGCCACGACGCAGTGCCCACAGGGACCACCAGGTGAACCTGGCCACCCTTGACTCCGAGGCCCT GCTGACCTTGGGCCTGAACCTCTCACACCTGGGCCGGGCCGAGCGCATCCTGGAGCTCCGGCCGGCCCTGGAGGGTCTAGA GGGCCGGATCCGCTACGTCATCGTCCGCGGAAACGAGCAAGGTTTCTTTCGCATGCATCACCTCCGTGGCGTCAGCTCCCT GCAGCTGGGGCGGAGGCGGCCGGGGCCTGGAACCTACCGGCTGGAGGTGGTGAGCCACATGGCAGGACCCTGGGGTGTCCA GCAAGAGGGGCAGCCAGGGCCATGGGGCCAGGCCTTGAGGCTGAAGGTGCAACTGTCAGTTGCTTTAGTTGGGAGGAGCCT CAGTGGGCCCCAGCTGTCCAGAGAAGGGGGATTCTGGAACTGGGAAGGACTGATCCCCAGAAGCGATGGCTGACCAGATTG AACCCCGAAACTCAGGAAGAGTGAAATGCTACACGACAACCTCAGGCAAGCCCGGCCTCTGCCTGGGCCTCTGTGCCAGCC CCGGGGGCCCCCCAGTTACTCAGTCTTTCCTGGAGACAGCAAGAAGCTGCAATGTGCAATCCCCCTGCCCCCACAGCCAAG GTCAGGAAGAGGCCCTGTGGTCACCGTGTCTGGCCAATCTCAGGCTTTCACTTCTGTACTGCACTGTGGCTTGCCCTGGCG GGGGGCAGGGGGTTGGCAGGACATGGCAATGGGCAACTGGGGTGGGCACAGGGCTTATTCCTCGGAGTAGAAGGGTGTACA GGGGGCCCAGACTCCACAGTGACTTGCCACATTTGCCCCCCATTTGGAGAATGCTTTTATATCAAAAGTGGAGACGATAAT AAAGTTATTTTGGGTTAAGTCTGCCTGCCCTTTGGCAAGTTCTTGAAGTAAGTAGATGCTGCCCTCGGACTGGGCGAGGCA GATCTTGTGCCTGGGGAAGCAGAAGGCCTTATGGGCTCCCCAGAATGGTAATAATGGCTCACGCTTCCTGACCACGTACTA CATACCAGACACCATTCGATTTTTTTTTTATTTTTTCTGAGACAGGGTCTTGCTCTGTTGCCTAGGTTGGAGTGCAGTGGC GCGATCGTGGCTCCCTGTGGTCGCCACTTTCCGGGCTCGAGCAATCCTCCCACCTCAGCCTCTCCCAAGTAGCTGGGACCA AAGGTGCACGCCACCACACCCAGCTAATTTTTTTAAATTTCTTTTTCTTTCTCTTTCTTTTTGAGACCAGCCCGGCCAACA TGACGAAACCCCATCTCTAATA

The disclosed NOV2d nucleic acid sequence, localized to chromsome 19, has 3194 of 4382 bases (72%) identical to a Mus musculus fibrillin 2 (fbn2) mRNA (gb:GENBANK- ID:MUSFBN2|acc:L39790.1) (E = 0.0).

A NOV2d polypeptide (SEQ ID NO: 12) encoded by SEQ ID NO:l 1 has 2845 amino acid residues and is presented using the one-letter code in Table 2H. Signal P, Psort and/or

Hydropathy results predict that NOV2d contains a signal peptide and is likely to be localized to the nucleus with a certainty of 0.6000. Although PSORT suggests that the Fibrillin-like protein may be localized in the nucleus, the NOV2d protein is similar to the Fibrillin family, some members of which are released extracellularly. Therefore it is likely that NOV2d protein shows a similar localization. The most likely cleavage site for a NOV2d peptide is between amino acids 29 and 30, at: AGG-QG. Table 2H. Encoded NOV2d protein sequence (SEQ ID NO:12).

MTLEGLY ARGPLAR LLAWSALLCMAGGQGRWDGA EAAGPGRVRRRGSPGILQGPNVCGSRFHAYCCPG RTFPGRSQC VVPICRRACGEGFCSQPNLCTCADGTLAPSCGVSRAICDRGCHNGGRCIGPNRCACVYGFMGPQCERDYRTGPCFGQVGPE GCQHQLTGLVCTKALCCATVGRAWGLPCELCPAQPHPCRRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCP VGHRLSDSSAACEDYDECSTIPGICEGGECTNTVSSYFCKCPPGFYTSPDGTLHGQSRAGACFSVLFGGRCAGDLAGHYTR RQCCCDRGRC AAGPVPELCPPRGSNEFQQLCAQR PLLPGHPGLFPGLLGFGSNGMGPP GPARLNPHGSDARGIPS GP GNSNIGTATLNQTIDICRHFTNLCLNGRCLPTPSSYRCECNVGYTQDVRGECIDVDECTSSPCHHGDCVNIPGTYHCRCYP GFQATPTRQACVDVDECIVSGG CHLGRCVNTEGSFQCVCNAGFELSPDGKNCVDHNECATST CVNGVCLNEDGSFSCLC KPGFLLAPGGHYCMDIDECQTPGICVNGHCTNTEGSFRCQCLGGLAVGTDGRVCVDTHVRSTCYGAIEKGSCARPFPGTVT KSECCCANPDHGFGEPCQLCPAKDSAEFQALCSSG GITTDGRDINECALDPEVCANGVCENLRGSYRCVCN GYEAGASG KDCTDVDECALNSLLCDNGWCQNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGS TCKCGPGSR DPS GTFCLDSTKGTCW KIQESRCEVNLQGASLRSECCATLGAAWGSPCERCEIDPACARGFARMTGVTCDDVNECESFPGVCP NGRCVNTAGSFRCECPEGL LDASGRLCVDVRLEPCFLRWDEDECGVT PGKYRMDVCCCSIGAVWGVECEACPDPESLEF ASLCPRG GFASRDFLSGRPFYKDVNECKVFPGLCTHGTCRNTVGSFHCACAGGFALDAQERNCTDIDECRISPD CGQGT CVNTPGSFECECFPGYESGFMLMKNCMDVDECARDPLLCRGGTCTNTDGSYKCQCPPGHELTAKGTACEDIDECSLSDGLC PHGQCVNVIGAFQCSCHAGFQSTPDRGATSASCPTEGHVQVVLGPGEQMCTG SIRAK STVPNPAPPDVDECEENPRVCD QGHCTNMPGGHRC CYDGFMATPDMRTCVDVDECD NPHICLHGDCENTKGSFVCHCQLGY VRKGATGCSDVDECEVGGH NCDSHASCLNIPGSFSCRCLPGWVGDGFECHD DECVSQEHRCSPRGDCLNVPGSYRCTCRQGFAGDGFFCEDRDECAENV DLCDNGQCLNAPGGYRCECEMGFDPTEDHRACQDVDECAQENLCAFGΞCEN PGMFRCICNGGYELDRGGGNCTDINECAD PVNCINGVCINTPGSYLCSCPQDFELNPSGVGCVDTRAGNCFLETHDRGDSGISCSAEIGVGVTRASCCCSLGRAWGNPCE LCPMANTTEYRTLCPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCECPPGYHLSEHTRICEDIDECSTHS GICGPGTCYNTLGNYTCVCPAEY QVNGGNNC D RKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCEACPTP ISPDYQI CGNQAPGFLTDIHTGKPLDIDECGEIPAICANGICINQIGSFRCECPAGFNYNSILLACEDVDECGSRESPCQ QNADCINIPGSYRCKCTRGYKLSPGGACVGRNECREIPNVCSHGDCMDTEGSYMCLCHRGFQASADQTLCMDIDECDRQPC GNGTCKNIIGSYNC CFPGFVVTHNGDCVDFDECTTLVGQVCRFGHC NTAGSFHC CQDGFELTADGKNCVDTNECLSLA GTCLPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLC FGTCTNSPGSFQCLCPPGFVLSDNGHRCFDTRQSFCF TRFEAGKCSVPKAFNTTKTRCCCSKRPGEGWGDPCELCPQEGSAAFQELCPFGHGAVPGPDDSREDVNECAENPGVCTNGV CVNTDGSFRCECPFGYS DFTGINCVDTDECSVGHPCGQGTCTNVIGGFECACADGFEPGLMMTCEDIDECSLNPLLCAFR CHNTEGSYLCTCPAGYTLREDGAMCRDVDECADGQQDCHARGMECKNLIGTFACVCPPGMRPLPGSGEGCTDDNECHAQPD LCVNGRCVNTAGSFRCDCDEGFQPSPTLTECHDIRQGPCFAEVLQTMCRΞLSSSSEAVTRAECCCGGGRGWGPRCELCPLP GTSAYRKLCPHGSGYTAEGRDVDECRMLAHLCAHGECINSLGSFRCHCQAGYTPDATATTCLDMDECSQVPKPCTFLCKNT KGSFLCSCPRGY LEEDGRTCKD DECTSRQHNCQFLCVNTVGAFTCRCPPGFTQHHQACFDNDECSAQPGPCGAHGHCHN TPGSFRCECHQGFTLVSSGHGCEDVNECDGPHRCQHGCQNQLGGYRCSCPQAFTQHSQWAQCVDENECALSPPTCGSASCR NTLGGFRCVCPSGFDFDQALGGCQEVDECAGRRGPCSYSCANTPGGFLCGCPQGYFRVGQGHCVSGLGFSPGPQDTPDKEE LLSSEACYECKINGLSPRDRPRRSAHRDHQVNLATLDSEALLT GLNLSH GRAERILELRPALEGLEGRIRYVIVRGNEQ GFFRMHHLRGVSS Q GRRRPGPGTYRLEVVSHMAGPWGVQQEGQPGPWGQA RLKVQLSVALVGRS SGPQLSREGGFWN WEGLIPRSDG

The NOV2d amino acid sequence has 1836 of 2753 amino acid residues (66%) identical to, and 2187 of 2753 amino acid residues (79%) similar to, a Mus musculus 2907 amino acid residue fibrillin 2 precursor protein (ptnr:SWISSPROT-ACC:Q61555) (E = 0.0).

NOV2d is expressed in at least the following tissues: adrenal gland, bone marrow, brain - amygdala, brain - cerebellum, brain - hippocampus, brain - substantia nigra, brain - thalamus, brain -whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma - Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea and uterus. Expression information was derived from the tissue sources ofthe sequences that were included in the derivation ofthe NOV2d sequence.

NOV2e

A disclosed NOV2e nucleic acid of 8219 nucleotides (also referred to as CG88987-03) encoding a novel Fibrillin-like protein is shown in Table 21. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 49-51 and ending with a TGA codon at nucleotides 8134-8136. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table 21, and the start and stop codons are in bold letters.

Table 21. NOV2e nucleotide sequence (SEQ ID NO:13).

ATCTAGGGCTGACTCTGCTTCCTCTCCGCAGCCTCCAGGGGACACGCCATGACTCTGGAGGGTCTGTATTTGGCAAGGGGC CCCCTGGCCCGGCTCCTGCTGGCCTGGTCGGCCCTGTTGTGCATGGCAGGTGGCCAAGGCCGCTGGGACGGGGCCTTGGAG GCTGCAGGTCCTGGACGTGTGCGGAGGCGGGGCAGCCCAGGCATCTTGCAGGGGCCGAATGTGTGCGGCTCCCGGTTCCAT GCCTACTGCTGTCCAGGCTGGAGGACATTCCCTGGCAGGAGCCAGTGTGTCGTACCCATCTGTAGGCGCGCCTGCGGTGAA GGCTTCTGCTCCCAGCCCAACCTGTGCACCTGTGCGGATGGGACGCTGGCTCCCAGCTGCGGGGTGAGCCGAGGGTCAGGG TGCAGTGTGAGCTGTATGAATGGGGGCACCTGCCGGGGGGCGTCCTGTCTGTGTCAGAAGGGCTACACAGGCACCGTGTGT GGGCAGCCCATCTGTGACCGCGGCTGCCACAATGGGGGTCGCTGCATTGGGCCCAACCGCTGCGCCTGTGTGTATGGCTTC ATGGGACCTCAATGTGAGAGAGATTACCGGACGGGATCCTGCTTTGGCCAAGTAGGCCCCGAGGGGTGCCAGCATCAGCTG ACGGGCCTCGTGTGCACCAAGGCACTTTGCTGTGCCACTGTGGGCCGTGCCTGGGGCCTTCCATGTGAACTTTGCCCTGCA CAGCCACACCCCTGCCGCCGCGGCTTCATCCCCAATATCCACACGGGGGCCTGCCAAGATGTGGATGAGTGCCAGGCTGTG CCAGGCCTGTGCCAGGGAGGCAGCTGCGTCAACATGGTGGGCTCCTTCCATTGCCGCTGTCCAGTTGGACACCGGCTCAGT GACAGCAGCGCCGCATGTGAAGACTACCGGGCCGGCGCCTGCTTCTCAGTGCTTTTCGGGGGCCGCTGTGCTGGAGACCTC GCCGGCCACTACACTCGCAGGCAGTGCTGCTGTGACAGGGGCAGGTGCTGGGCAGCTGGCCCGGTCCCTGAGCTGTGTCCT CCTCGGGGCTCCAATGAATTCCAGCAACTGTGCGCCCAGCGGCTGCCGCTGCTACCCGGCCACCCTGGCCTCTTCCCTGGC CTCCTGGGCTTCGGATCCAATGGCATGGGTCCCCCTCTTGGGCCAGCGCGACTCAACCCCCATGGCTCTGATGCGCGTGGG ATCCCCAGCCTGGGCCCTGGCAACTCTAATATTGGCACTGCTACCCTGAACCAGACCATTGACATCTGCCGACACTTCACC AACCTGTGTCTGAATGGCCGCTGCCTGCCCACGCCTTCCAGCTACCGCTGCGAGTGTAACGTGGGCTACACCCAGGACGTG CGCGGCGAGTGCATTGATGTAGACGAATGCACCAGCAGCCCCTGCCACCACGGTGACTGCGTCAACATCCCCGGCACCTAC CACTGCCGGTGCTACCCGGGCTTCCAGGCCACGCCCACCAGGCAGGCATGCGTGGATGTGGACGAGTGCATTGTCAGTGGT GGCCTTTGTCACCTGGGCCGCTGTGTCAACACAGAGGGCAGCTTCCAGTGTGTCTGCAATGCAGGCTTCGAGCTCAGCCCT GACGGCAAGAACTGTGTGGACCACAACGAGTGTGCCACCAGCACCATGTGCGTCAACGGCGTGTGTCTCAACGAGGATGGC AGCTTCTCCTGCCTCTGCAAACCCGGCTTCCTGCTGGCGCCTGGCGGCCACTACTGCATGGACATTGACGAGTGCCAGACG CCCGGCATCTGCGTGAACGGCCACTGTACCAACACCGAGGGCTCCTTCCGCTGCCAGTGCCTGGGGGGGCTGGCGGTAGGC ACGGATGGCCGCGTGTGCGTGGACACCCACGTGCGCAGCACCTGCTATGGGGCCATCGAGAAGGGCTCCTGTGCCCGCCCC TTCCCTGGCACTGTCACCAAGTCGGAGTGCTGCTGTGCCAATCCGGACCACGGTTTTGGGGAGCCCTGCCAGCTTTGTCCT GCCAAAAACTCCGCTGAGTTCCAGGCACTGTGCAGCAGTGGGCTTGGCATTACCACGGATGGTCGAGACATCAACGAGTGT GCTCTGGATCCTGAGGTTTGTGCCAATGGCGTGTGCGAGAACCTTCGGGGCAGCTACCGCTGTGTCTGCAACCTGGGTTAT GAGGCAGGTGCCTCAGGCAAGGACTGCACAGACGTGGATGAGTGTGCCCTCAACAGCCTCCTGTGTGACAACGGGTGGTGC CAGAATAGCCCTGGCAGCTACAGCTGCTCCTGCCCCCCCGGCTTCCACTTCTGGCAGGACACGGAGATCTGCAAAGATGTC GACGAATGCCTGTCCAGCCCGTGTGTGAGTGGCGTTTGTCGGAACCTGGCCGGCTCCTACACCTGCAAATGTGGCCCTGGC AGCCGGCTGGACCCCTCTGGTACCTTCTGTCTAGACAGCACCAAGGGCACCTGCTGGCTGAAGATCCAGGAGAGCCGCTGT GAGGTGAACCTTCAGGGAGCCAGCCTGCGGTCTGAGTGCTGCGCCACCCTCGGGGCAGCCTGGGGGAGCCCCTGCGAACGC TGCGAGATCGACCCTGCCTGTGCCCGGGGCTTTGCCCGGATGACGGGTGTCACCTGCGATGATGTGAACGAGTGTGAGTCC TTCCCGGGAGTCTGTCCCAACGGGCGTTGCGTCAACACTGCTGGGTCTTTCCGCTGTGAGTGTCCAGAGGGCCTGATGCTG GACGCCTCAGGCCGGCTGTGCGTGGATGTGAGATTGGAACCATGTTTCCTGCGATGGGATGAGGATGAGTGTGCAAGGGAC CCGCTGCTCTGCTGGGGAGGCACTTGCACCAACACGGATGGGAGCTACAAGTGCCAGTGTCCCCCTGGGCATGAGCTGACG GCCAAGGGCACTGCCTGTGAGGACATCGATGAGTGCTCCCTGAGTGATGGCCTGTGTCCCCATGGCCAGTGTGTCAATGTC ATCGGTGCCTTCCAGTGCTCCTGCCATGCCGGCTTCCAGAGCACACCTGACCGCCAGGGCTGCGTGGACATCAACGAATGC CGGGTCCAGAATGGTGGGTGTGACGTGCACTGTATTAACACTGAGGGCAGCTACCGGTGCAGCTGTGGGCAGGGCTACTCG CTGATGCCCGACGGAAGGGCATGTGCAGACGTGGACGAGTGTGAAGAGAACCCCCGCGTTTGTGACCAAGGCCACTGCACC AACATGCCAGGGGGTCACCGCTGCCTGTGCTATGATGGCTTCATGGCCACGCCAGACATGAGGACATGTGTTGATGTGGAT GAGTGTGACCTGAACCCTCACATCTGCCTCCATGGGGACTGCGAGAACACGAAGGGTTCCTTTGTCTGCCACTGTCAGCTG GGCTACATGGTCAGGAAGGGGGCCACAGGCTGCTCTGATGTGGATGAATGCGAGGTTGGAGGACACAACTGTGACAGTCAC GCCTCCTGTCTCAACATCCCGGGGAGTTTCAGCTGTAGGTGCCTGCCAGGCTGGGTGGGGGATGGCTTCGAATGTCACGAC CTGGATGAATGCGTCTCCCAGGAGCACCGGTGCAGCCCAAGAGGTGACTGTCTCAATGTCCCTGGCTCCTACCGCTGCACC TGCCGCCAGGGCTTTGCCGGGGATGGCTTCTTCTGCTCAGACAGGGATGAATGTGCCGAGAACGTGGACCTCTGTGACAAC GGGCAGTGCCTCAATGCGCCCGGCGGGTACCGCTGTGAATGTGAGATGGGCTTTGACCCCACCGAGGACCACCGGGCCTGC CAGATGGACGAGTGTGCGCAAGGGAACCTCTGTGCATTTGGGAGCTGTGAGAACCTGCCTGGAATGTTCCGCTGCATCTGC AATGGTGGCTACGAACTGGACCGAGGGGGTGGCAACTGCACAACAGACATCAACGAGTGTGCAGACCCAGTAAACTGCATC AACGGCGTGTGCATTAACACCCCCGGCAGCTACCTCTGCAGCTGCCCCCAGGATTTTGAGCTGAACCCCAGCGGAGTGGGC TGCGTGGACACTCGGGCCGGGAACTGTTTCCTGGAGACGCATGACCGAGGGGACAGTGGCATTTCCTGCAGTGCCGAGATC GGAGTTGGTGTCACCCGAGCTTCCTGCTGTTGCTCCCTGGGCCGGGCTTGGGGCAATCCCTGTGAGCTGTGCCCTATGGCC AACACCACTGAGTACAGAACCCTGTGCCCGGGTGGTGAGGGCTTCCAGCCTAACCGCATCACTGTCATTCTGGAAGACATC GACGAGTGCCAAGAGCTGCCAGGGCTGTGTCAGGGGGGTGACTGCGTCAACACGTTTGGCAGTTTCCAGTGTGAGTGCCCA CCTGGCTACCACCTCAGTGAGCACACCCGCATCTGTGAGGATATTGACGAATGCTCCACACACTCCGGCATCTGTGGCCCT GGCACCTGCTACAACACCCTGGGGAACTACACCTGTGTCTGCCCTGCAGAGTACCTCCAAGTCAATGGTGGCAACAACTGC ATGGATATGAGGAAGAGTGTCTGCTTCCGGCACTATAACGGCACATGTCAAAATGAGCTGGCCTTCAACGTGACCCGGAAA ATGTGTTGCTGCTCCTACAACATTGGCCAGGCCTGGAATAGACCCTGTGAGGCCTGCCCCACTCCCATCAGTCCTGACTAC CAGATCCTGTGTGGAAATCAGGCCCCGGGATTCCTCACTGACATCCACACGGGGAAGCCCCTTGACATTGATGAGTGTGGG GAGATCCCCGCCATCTGTGCCAATGGCATCTGCATAAACCAGATCGGGAGTTTCCGCTGCGAGTGCCCCGCAGGCTTCAAC TACAACAGCATCCTGCTGGCTTGTGAAGATGTCGATGAGTGTGGCAGCAGGGAGAGTCCCTGCCAGCAGAATGCTGACTGC ATCAACATCCCCGGTAGCTACCGCTGCAAGTGCACCCGAGGGTACAAACTGTCGCCAGGCGGGGCTTGTGTGGGACGGAAT GAGTGTCGGGAGATCCCGAATGTCTGTAGCCATGGTGACTGCATGGACACAGAAGGCAGCTACATGTGTCTGTGTCACCGT GGATTCCAGGCCTCTGCAGACCAGACCCTGTGCATGGACATTGACGAGTGTGACCGGCAGCCTTGTGGAAATGGGACCTGC AAGAACATCATTGGCTCCTACAACTGCCTCTGCTTCCCTGGCTTTGTGGTGACACACAATGGGGATTGTGTGGATTTTGAT GAGTGTACTACCCTGGTGGGGCAGGTGTGCCGATTTGGCCATTGCCTCAACACAGCTGGTTCCTTCCACTGCCTCTGCCAG GATGGCTTTGAGCTCACAGCTGATGGGAAGAACTGTGTGGACACCAATGAGTGCCTCAGCCTTGCAGGAACCTGCCTACCC GGCACTTGCCAGAACCTCGAGGGCTCCTTCCGCTGCATCTGTCCCCCTGGCTTCCAGGTGCAGAGTGACCACTGCATTGAT ATCGACGAGTGCTCAGAGGAGCCCAACCTCTGCCTCTTTGGCACCTGTACCAACAGCCCTGGGAGCTTCCAGTGCCTCTGC CCACCTGGCTTTGTCCTCTCTGACAATGGGCACCGTTGCTTTGACACACGGCAGAGTTTCTGCTTCACCCGTTTTGAGGCT GGGAAGTGCTCGGTGCCCAAAGCTTTCAACACCACCAAGACCCGCTGCTGCTGCAGTAAGAGGCCTGGGGAGGGCTGGGGA GACCCCTGCGAACTGTGTCCCCAGGAGGACTCCCCTCCCCCTCTCCGTCCAGCTGCCTTTCAGGAGCTCTGCCCCTTTGGC CACGGGGCAGTCCCAGGCCCGGATGACTCCCGAGAAGACGTGAATGAGTGTGCAGAGAACCCTGGCGTCTGCACTAACGGC GTCTGTGTCAACACCGATGGATCCTTCCGCTGTGAGTGTCCCTTTGGCTACAGCCTGGACTTCACTGGCATCAACTGTGAG GACACAGACGAGTGCTCTGTCGGCCACCCCTGTGGGCAAGGGACATGCACCAATGTCATCGGAGGCTTCGAATGTGCCTGT GCTGACGGCTTTGAGCCTGGCCTCATGATGACCTGCGAGGACATCGACGAATGCTCCCTGAACCCGCTGCTCTGTGCCTTC CGCTGCCACAATACCGAGGGCTCCTACCTGTGCACCTGTCCAGCCGGCTACACCCTGCGGGAGGATGGGGCCATGTGTCGA GATGTGGACGAGTGTGCAGATGGTCAGCAGGACTGCCACGCCCGGGGCATGGAGTGCAAGAACCTCATCGGTACCTTCGCG TGCGTCTGTCCCCCAGGCATGCGGCCCCTGCCTGGCTCTGGGGAGGGCTGCACAGATGACAATGAATGCCACGCTCAGCCT GACCTCTGTGTCAACGGCCGCTGTGTCAACACCGCGGGCAGCTTCCGGTGCGACTGTGATGAGGGATTCCAGCCCAGCCCC ACCCTTACCGAGTGCCACGACATCCGGCAGGGGCCCTGCTTTGCCGAGGTGCTGCAGACCATGTGCCGGTCTCTGTCCAGC AGCAGTGAGGCTGTCACCAGGGCCGAGTGCTGCTGTGGGGGTGGCCGGGGCTGGGGGCCCCGCTGCGAGCTCTGTCCCCTG CCCGGCACCTCTGCCTACAGGAAGCTGTGCCCCCATGGCTCAGGCTACACTGCTGAGGGCCGAGATGTAGATGAATGCCGT ATGCTTGCTCACCTGTGTGCTCATGGGGAGTGCATCAACAGCCTTGGCTCCTTCCGCTGCCACTGTCAGGCCGGGTACACA CCGGATGCTACTGCTACTACCTGCCTGGATATGGATGAGTGCAGCCAGGTCCCCAAGCCATGTACCTTCCTCTGCAAAAAC ACGAAGGGCAGTTTCCTGTGCAGCTGTCCCCGAGGCTACCTGCTGGAGGAGGATGGCAGGACCTGCAAAGACCTGGACGAA TGCACCTCCCGGCAGCACAACTGTCAGTTCCTCTGTGTCAACACTGTGGGCGCCTTCACCTGCCGCTGTCCACCCGGCTTC ACCCAGCACCACCAGGCCTGCTTCGACAATGATGAGTGCTCAGCCCAGCCTGGCCCATGTGGTGCCCACGGGCACTGCCAC AACACCCCGGGCAGCTTCCGCTGTGAATGCCACCAAGGCTTCACCCTGGTCAGCTCAGGCCATGGCTGTGAAGATGTGAAT GAATGTGATGGGCCCCACCGCTGCCAGCATGGCTGTCAGAACCAGCTAGGGGGCTACCGCTGCAGCTGCCCCCAGGCTTTC ACCCAGCACTCCCAGTGGGCCCAGTGTGTGGATGAGAATGAGTGTGCCCTGTCGCCCCCCACCTGCGGGAGCGCCTCCTGT CGCAACACTCTTGGTGGCTTCCGCTGCGTCTGCCCCTCCGGCTTTGACTTTGATCAGGCCCTCGGGGGCTGCCAGGAGGTG GATGAGTGCGCCGGACGGCGTGGCCCCTGTAGCTACAGCTGTGCCAACACGCCTGGTGGCTTCCTGTGCGGCTGTCCTCAA GGCTACTTCCGGGTTGGGCAAGGGCACTGTGTCTCCGGCCTGGGCTTCAGCCCCGGACCCCAGGACACCCCGGACAAAGAG GAGCTGCTCTCGTCTGAAGCCTGCTACGAATGCAAGATCAATGGCCTCTCCCCTCGGGACCGGCCACGACGCAGTGCCCAC AGGGACCACCAGGTGAACCTGGCCACCCTTGACTCCGAGGCCCTGCTGACCTTGGGCCTGAACCTCTCACACCTGGGCCGG GCCGAGCGCATCCTGGAGCTCCGGCCGGCCCTGGAGGGTCTAGAGGGCCGGATCCGCTACGTCATCGTCCGCGGAAACGAG CAAGGTTTCTTTCGCATGCATCACCTCCGTGGCGTCAGCTCCCTGCAGCTGGGGCGGAGGCGGCCGGGGCCTGGAACCTAC CGGCTGGAGGTGGTGAGCCACATGGCAGGACCCTGGGGTGTCCAGCAAGAGGGGCAGCCAGGGCCATGGGGCCAGGCCTTG AGGCTGAAGGTGCAACTGTCAGTTGCTTTAGTTGGGAGGAGCCTCAGTGGGCCCCAGCTGTCCAGAGAAGGGGGATTCTGG AACTGGGAAGGACTGATCCCCAGAAGCGATGGCTGACCAGATTGAACCCCGAAACTCAGGAAGAGTGAAATGCTACACGAC AACCTCAGGCAAGCCCGGCCTCTGCCTGGGCCTCTGTG

The disclosed NOV2e nucleic acid sequence, localized to chromsome 19, has 2166 of 2977 bases (72%) identical to a Mus musculus fibrillin 2 (fbn2) mRNA (gb:GENBANK- ID:MUSFBN2|acc:L39790.1) (E = 0.0).

A NOV2e polypeptide (SEQ ID NO: 14) encoded by SEQ ID NO: 13 has 2695 amino acid residues and is presented using the one-letter code in Table 2J. Signal P, Psort and/or Hydropathy results predict that NOV2e contains a signal peptide and is likely to be localized to the nucleus with a certainty of 0.6000. Although PSORT suggests that the Fibrillin-like protein may be localized in the nucleus, the NOV2e protein is similar to the Fibrillin family, some members of which are released extracellularly. Therefore it is likely that NOV2e protein shows a similar localization. The most likely cleavage site for a NOV2e peptide is between amino acids 29 and 30, at: AGG-QG.

Table 2J. Encoded NOV2e protein sequence (SEQ ID NO:14).

MTLEGLY ARGPLAR L AWSAL C AGGQGRWDGALEAAGPGRVRRRGSPGILQGPNVCGSRFHAYCCPGWRTFPGRSQC VVPICRRACGEGFCSQPNLCTCADGTLAPSCGVSRGSGCSVSCMNGGTCRGASCLCQKGYTGTVCGQPICDRGCHNGGRCI GPNRCACVYGFMGPQCERDYRTGSCFGQVGPEGCQHQLTGLVCTKALCCATVGRAWGLPCELCPAQPHPCRRGFIPNIHTG ACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVGHRLSDSSAACEDYRAGACFSV FGGRCAGD AGHYTRRQCCCDRGRC WAAGPVPELCPPRGSNEFQQLCAQR PL PGHPGLFPGLLGFGSNGMGPP GPAR NPHGSDARGIPSLGPGNSNIGTATL NQTIDICRHFTNLCLNGRCLPTPSSYRCECNVGYTQDVRGECIDVDECTSSPCHHGDCVNIPGTYHCRCYPGFQATPTRQA CVDVDECIVSGGLCHLGRCVNTEGSFQCVCNAGFELSPDGKNCVDHNECATSTMCVNGVCLNEDGSFSCLCKPGFLLAPGG HYCMDIDECQTPGICVNGHCTNTEGSFRCQCLGG AVGTDGRVCVDTHVRSTCYGAIEKGΞCARPFPGTVTKSECCCANPD HGFGEPCQLCPAKNSAEFQALCSSGLGITTDGRDINECA DPEVCANGVCEN RGSYRCVCNLGYEAGASGKDCTDVDECA LNSL CDNGWCQNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGSYTCKCGPGSRLDPSGTFCLDSTKG TCWLKIQESRCEVN QGASLRSECCATLGAAWGSPCERCEIDPACARGFARMTGVTCDDVNECESFPGVCPNGRCVNTAGS FRCECPEGLMLDASGRLCVDVRLEPCFLRWDEDECARDPLLCWGGTCTNTDGSYKCQCPPGHELTAKGTACEDIDECS SD G CPHGQCVNVIGAFQCSCHAGFQSTPDRQGCVDINECRVQNGGCDVHCINTEGSYRCSCGQGYSLMPDGRACADVDECEE O 02 0

NPRVCD_QGHCTNMPGGHRC CYDGFMATPDMRTCVDVDECDLNPHICLHGDCENTKGSFVCHCQLGYMVRKGATGCSDVDE CEVGGHNCDSHASC NIPGSFSCRCLPGWVGDGFECHD DECVSQEHRCSPRGDCLNVPGSYRCTCRQGFAGDGFFCSDRD ECAENVDLCDNG_QCLNAPGGYRCECEMGFDPTEDHRACQMDECAQGNLCAFGSCENLPG FRCICNGGYELDRGGGNCTTD INECADPVNCINGVCINTPGSYLCSCP_QDFE NPSGVGCVDTRAGNCFLETHDRGDSGISCSAEIGVGVTRASCCCS GRA GNPCELCPMANTTEYRTLCPGGEGF_QPNRITVILEDIDECQELPG CQGGDCVNTFGSFQCECPPGYHLSEHTRICEDID ECSTHSGICGPGTCYNTLGNYTCVCPAEYLQVNGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQA NRPC EACPTPISPDYQI CGNQAPGF TDIHTGKPLDIDECGEIPAICANGICINQIGSFRCECPAGFNYNSI LACEDVDECGS RESPC_QQNADCINIPGSYRCKCTRGYKLSPGGACVGRNECREIPNVCSHGDC DTEGSYMCLCHRGFQASADQTLCMDIDE CDRQPCGNGTCKNIIGSYNCLCFPGFVVTHNGDCVDFDECTTLVGQVCRFGHC NTAGSFHCLCQDGFELTADGKNCVDTN EC SLAGTC PGTCQNLEGSFRCICPPGF_QVQSDHCIDIDECSEEPNLCLFGTCTNSPGSFQC CPPGFVLSDNGHRCFDT R_QSFCFTRFEAGKCSVPKAFNTTKTRCCCSKRPGEGWGDPCELCPQEDSPPPLRPAAFQE CPFGHGAVPGPDDSREDVNE CAENPGVCTNGVCVNTDGSFRCECPFGYSLDFTGINCEDTDECSVGHPCGQGTCTNVIGGFECACADGFEPGLMMTCEDID ECSLNPLLCAFRCHNTEGSYLCTCPAGYTLREDGAMCRDVDECADGQQDCHARGMECKNLIGTFACVCPPGMRPLPGSGEG CTDDNECHA_QPDLCVNGRCVNTAGSFRCDCDEGFQPSPTLTECHDIRQGPCFAEV QTMCRSLΞSSSEAVTRAECCCGGGR G GPRCELCPLPGTSAYRK CPHGSGYTAEGRDVDECRMLAHLCAHGECINS GSFRCHCQAGYTPDATATTCLDMDECSQ VPKPCTFLCKNTKGSFLCSCPRGYLLEEDGRTCKDLDECTSRQHNCQF CVNTVGAFTCRCPPGFTQHHQACFDNDECSAQ PGPCGAHGHCHNTPGSFRCECH_QGFTLVSSGHGCEDVNECDGPHRCQHGCQNQLGGYRCSCPQAFTQHSQWAQCVDENECA LSPPTCGSASCRNTLGGFRCVCPSGFDFD_QALGGCQEVDECAGRRGPCSYSCANTPGGFLCGCPQGYFRVGQGHCVSGLGF ΞPGP_QDTPDKEELLSSEACYECKINGLSPRDRPRRSAHRDHQVNLATLDSEAL TLG NLSHLGRAERILE RPALEGLEG RIRYVIVRGNE_QGFFRMHHLRGVSS QLGRRRPGPGTYRLEVVSHMAGPWGVQQEGQPGPWGQALR KVQLSVA VGRSLS GPQLSREGGFWNWEG IPRSDG

The NOV2e amino acid sequence has 1881 of 2256 amino acid residues (83%) identical to, and 1935 of 2256 amino acid residues (85%) similar to, a Homo sapiens 2809 amino acid residue fibrillin 3 protein (ptnr:TREMBLNEW-ACC:BAB47408) (E = 0.0). NOV2e is expressed in at least the following tissues: lung, colon, bone, trabecular bone cells, placenta, germ cell, melanocyte, heart, uterus, thyroid and brain. Expression information was derived from the tissue sources ofthe sequences that were included in the derivation of the NOV2e sequence.

NOV2f A disclosed NOV2f nucleic acid of 9154 nucleotides (also referred to as CG88987-05) encoding a novel Fibrillin-like protein is shown in Table 2K. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 41-43 and ending with a TGA codon at nucleotides 8312-8314. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table 2K, and the start and stop codons are in bold letters.

Table 2K. NO V2f nucleotide sequence (SEQ ID NO:15).

CTGACTCTGCTTCCTCTCCGCAGCCTCCAGGGGACACGCCATGACTCTGGAGGGTCTGTATTTGGCAAGGGGCCCCCTGGC CCGGCTCCTGCTGGCCTGGTCGGCCCTGTTGTGCATGGCAGGTGGCCAAGGCCGCTGGGACGGGGCCTTGGAGGCTGCAGG TCCTGGACGTGTGCGGAGGCGGGGCAGCCCAGGCATCTTGCAGGGGCCGAATGTGTGCGGCTCCCGGTTCCATGCCTACTG CTGTCCAGGCTGGAGGACATTCCCTGGCAGGAGCCAGTGTGTCGTACCCATCTGTAGGCGCGCCTGCGGTGAAGGCTTCTG CTCCCAGCCCAACCTGTGCACCTGTGCGGATGGGACGCTGGCTCCCAGCTGCGGGGTGAGCCGAGCCATCTGTGACCGCGG CTGCCACAATGGGGGTCGCTGCATTGGGCCCAACCGCTGCGCCTGTGTGTATGGCTTCATGGGACCTCAATGTGAGAGAGA TTACCGGACGGGACCCTGCTTTGGCCAAGTAGGCCCCGAGGGGTGCCAGCATCAGCTGACGGGCCTCGTGTGCACCAAGGC ACTTTGCTGTGCCACTGTGGGCCGTGCCTGGGGCCTTCCATGTGAACTTTGCCCTGCACAGCCACACCCCTGCCGCCGCGG CTTCATCCCCAATATCCACACGGGGGCCTGCCAAGATGTGGATGAGTGCCAGGCTGTGCCAGGCCTGTGCCAGGGAGGCAG CTGCGTCAACATGGTGGGCTCCTTCCATTGCCGCTGTCCAGTTGGACACCGGCTCAGTGACAGCAGCGCCGCATGTGAAGA CTACGATGAATGCAGCACCATTCCTGGAATCTGTGAAGGGGGTGAATGTACAAACACAGTCAGCAGTTACTTTTGCAAATG TCCTCCTGGTTTTTACACCTCTCCAGATGGTACTCTTCATGGACAGTCGCGGGCCGGCGCCTGCTTCTCAGTGCTTTTCGG GGGCCGCTGTGCTGGAGACCTCGCCGGCCACTACACTCGCAGGCAGTGCTGCTGTGACAGGGGCAGGTGCTGGGCAGCTGG CCCGGTCCCTGAGCTGTGTCCTCCTCGGGGCTCCAATGAATTCCAGCAACTGTGCGCCCAGCGGCTGCCGCTGCTACCCGG CCACCCTGGCCTCTTCCCTGGCCTCCTGGGCTTCGGATCCAATGGCATGGGTCCCCCTCTTGGGCCAGCGCGACTCAACCC CCATGGCTCTGATGCGCGTGGGATCCCCAGCCTGGGCCCTGGCAACTCTAATATTGGCACTGCTACCCTGAACCAGACCAT TGACATCTGCCGACACTTCACCAACCTGTGTCTGAATGGCCGCTGCCTGCCCACGCCTTCCAGCTACCGCTGCGAGTGTAA CGTGGGCTACACCCAGGACGTGCGCGGCGAGTGCATTGATGTAGACGAATGCACCAGCAGCCCCTGCCACCACGGTGACTG CGTCAACATCCCCGGCACCTACCACTGCCGGTGCTACCCGGGCTTCCAGGCCACGCCCACCAGGCAGGCATGCGTGGATGT GGACGAGTGCATTGTCAGTGGTGGCCTTTGTCACCTGGGCCGCTGTGTCAACACAGAGGGCAGCTTCCAGTGTGTCTGCAA TGCAGGCTTCGAGCTCAGCCCTGACGGCAAGAACTGTGTGGACCACAACGAGTGTGCCACCAGCACCATGTGCGTCAACGG CGTGTGTCTCAACGAGGATGGCAGCTTCTCCTGCCTCTGCAAACCCGGCTTCCTGCTGGCGCCTGGCGGCCACTACTGCAT GGACATTGACGAGTGCCAGACGCCCGGCATCTGCGTGAACGGCCACTGTACCAACACCGAGGGCTCCTTCCGCTGCCAGTG CCTGGGGGGGCTGGCGGTAGGCACGGATGGCCGCGTGTGCGTGGACACCCACGTGCGCAGCACCTGCTATGGGGCCATCGA GAAGGGCTCCTGTGCCCGCCCCTTCCCTGGCACTGTCACCAAGTCCGAGTGCTGCTGTGCCAATCCGGACCACGGTTTTGG GGAGCCCTGCCAGCTTTGTCCTGCCAAAGACTCCGCTGAGTTCCAGGCACTGTGCAGCAGTGGGCTTGGCATTACCACGGA TGGTCGAGACATCAACGAGTGTGCTCTGGATCCTGAGGTTTGTGCCAATGGCGTGTGCGAGAACCTTCGGGGCAGCTACCG CTGTGTCTGCAACCTGGGTTATGAGGCAGGTGCCTCAGGCAAGGACTGCACAGACGTGGATGAGTGTGCCCTCAACAGCCT CCTGTGTGACAACGGGTGGTGCCAGAATAGCCCTGGCAGCTACAGCTGCTCCTGCCCCCCCGGCTTCCACTTCTGGCAGGA CACGGAGATCTGCAAAGATGTCGACGAATGCCTGTCCAGCCCGTGTGTGAGTGGCGTCTGTCGGAACCTGGCCGGCTCCTA CACCTGCAAATGTGGCCCTGGCAGCCGGCTGGACCCCTCTGGTACCTTCTGTCTAGACAGCACCAAGGGCACCTGCTGGCT GAAGATCCAGGAGAGCCGCTGTGAGGTGAACCTTCAGGGAGCCAGCCTGCGGTCTGAGTGCTGTGCCACCCTCGGGGCAGC CTGGGGGAGCCCCTGCGAACGCTGCGAGATCGACCCTGCCTGTGCCCGGGGCTTTGCCCGGATGACGGGTGTCACCTGCGA TGATGTGAACGAGTGTGAGTCCTTCCCGGGAGTCTGTCCCAACGGGCGTTGCGTCAACACTGCTGGGTCTTTCCGCTGTGA GTGTCCAGAGGGCCTGATGCTGGACGCCTCAGGCCGGCTGTGCGTGGATGTGAGATTGGAACCATGTTTCCTGCGATGGGA TGAGGATGAGTGTGGGGTCACCCTGCCTGGCAAGTACCGGATGGACGTCTGCTGCTGCTCCATCGGGGCCGTGTGGGGAGT CGAGTGCGAGGCCTGCCCGGATCCCGAGTCTCTGGAGTTCGCCAGCCTGTGCCCGCGGGGGCTGGGCTTCGCCAGCCGGGA CTTCCTGTCTGGCCGACCATTCTATAAAGATGTGAATGAATGCAAGGTGTTCCCTGGCCTCTGCACGCACGGTACCTGCAG AAACACGGTGGGCAGCTTCCACTGCGCCTGTGCGGGGGGCTTCGCCCTGGATGCCCAGGAACGGAACTGCACAGATATCGA CGAGTGTCGCATCTCTCCTGACCTCTGCGGCCAGGGCACCTGTGTCAACACGCCGGGCAGCTTTGAGTGCGAGTGTTTTCC CGGCTACGAGAGTGGCTTCATGCTGATGAAGAACTGCATGGACGTGGACGAGTGTGCAAGGGACCCGCTGCTCTGCCGGGG AGGCACTTGCACCAACACGGATGGGAGCTACAAGTGCCAGTGTCCCCCTGGGCATGAGCTGACGGCCAAGGGCACTGCCTG TGAGGACATCGATGAGTGCTCCCTGAGTGATGGCCTGTGTCCCCATGGCCAGTGTGTCAATGTCATCGGTGCCTTCCAGTG CTCCTGCCATGCCGGCTTCCAGAGCACACCTGACCGCGGAGCTACAAGTGCCAGTTGCCCGACGGAAGGGCATGTGCAGGT CGTCCTGGGGCCTGGGGAGCAGATGTGTACTGGGTGGTCTATCAGGGCAAAGCTAAGCACAGTCCCCAACCCTGCTCCCCC AGACGTGGACGAGTGTGAAGAGAACCCCCGCGTTTGTGACCAAGGCCACTGCACCAACATGCCAGGGGGTCACCGCTGCCT GTGCTATGATGGCTTCATGGCCACGCCAGACATGAGGACATGTGTTGATGTGGATGAGTGTGACCTGAACCCTCACATCTG CCTCCATGGGGACTGCGAGAACACGAAGGGTTCCTTTGTCTGCCACTGTCAGCTGGGCTACATGGTCAGGAAGGGGGCCAC AGGCTGCTCTGATGTGGATGAATGCGAGGTTGGAGGACACAACTGTGACAGTCACGCCTCCTGTCTCAACATCCCAGGGAG TTTCAGCTGTAGGTGCCTGCCAGGCTGGGTGGGGGATGGCTTCGAATGTCACGACCTGGATGAATGCGTCTCCCAGGAGCA CCGGTGCAGCCCAAGAGGTGACTGTCTCAATGTCCCTGGCTCCTACCGCTGCACCTGCCGCCAGGGCTTTGCCGGGGATGG CTTCTTCTGCGAAGACAGGGATGAATGTGCCGAGAACGTGGACCTCTGTGACAACGGGCAGTGCCTCAATGCGCCCGGCGG GTACCGCTGTGAATGTGAGATGGGCTTTGACCCCACCGAGGACCACCGGGCCTGCCAGGATGTGGACGAGTGTGCGCAAGA GAACCTCTGTGCATTTGGGAGCTGTGAGAACCTGCCTGGAATGTTCCGCTGCATCTGCAATGGTGGCTACGAACTGGACCG AGGGGGTGGCAACTGCACAGACATCAACGAGTGTGCAGACCCAGTAAACTGCATCAACGGCGTGTGCATTAACACCCCCGG CAGCTACCTCTGCAGCTGCCCCCAGGATTTTGAGCTGAACCCCAGCGGAGTGGGCTGCGTGGACACTCGGGCCGGGAACTG TTTCCTGGAGACGCATGACCGAGGGGACAGTGGCATTTCCTGCAGTGCCGAGATCGGAGTTGGTGTCACCCGAGCTTCCTG CTGTTGCTCCCTGGGCCGGGCTTGGGGCAATCCCTGTGAGCTGTGCCCTATGGCCAACACCACTGAGTACAGAACCCTGTG CCCGGGTGGTGAGGGCTTCCAGCCTAACCGCATCACTGTCATTCTGGAAGACATCGACGAGTGCCAAGAGCTGCCAGGGCT GTGTCAGGGGGGTGACTGCGTCAACACGTTTGGCAGTTTCCAGTGTGAGTGCCCACCTGGCTACCACCTCAGTGAGCACAC CCGCATCTGTGAGGATATTGACGAATGCTCCACACACTCCGGCATCTGTGGCCCTGGCACCTGCTACAACACCCTGGGGAA CTACACCTGTGTCTGCCCTGCAGAGTACCTCCAAGTCAATGGTGGCAACAACTGCATGGATATGAGGAAGAGTGTCTGCTT CCGGCACTATAACGGCACATGTCAAAATGAGCTGGCCTTCAACGTGACCCGGAAAATGTGTTGCTGCTCCTACAACATTGG CCAGGCCTGGAATAGACCCTGTGAGGCCTGCCCCACTCCCATCAGTCCTGACTACCAGATCCTGTGTGGAAATCAGGCCCC GGGATTCCTCACTGACATCCACACGGGGAAGCCCCTTGACATTGATGAGTGTGGGGAGATCCCCGCCATCTGTGCCAATGG CATCTGCATAAACCAGATCGGGAGTTTCCGCTGCGAGTGCCCCGCAGGCTTCAACTACAACAGCATCCTGCTGGCTTGTGA AGATGTCGATGAGTGTGGCAGCAGGGAGAGTCCCTGCCAGCAGAATGCTGACTGCATCAACATCCCCGGTAGCTACCGCTG CAAGTGCACCCGAGGGTACAAACTGTCGCCAGGCGGGGCTTGTGTGGGACGGAATGAGTGTCGGGAGATCCCGAATGTCTG TAGCCATGGTGACTGCATGGACACAGAAGGCAGCTACATGTGTCTGTGTCACCGTGGATTCCAGGCCTCTGCAGACCAGAC CCTGTGCATGGACATTGACGAGTGTGACCGGCAGCCTTGTGGAAATGGGACCTGCAAGAACATCATTGGCTCCTACAACTG CCTCTGCTTCCCTGGCTTTGTGGTGACACACAATGGGGATTGTGTGGATTTTGATGAGTGTACTACCCTGGTGGGGCAGGT GTGCCGATTTGGCCATTGCCTCAACACAGCTGGTTCCTTCCACTGCCTCTGCCAGGATGGCTTTGAGCTCACAGCTGATGG GAAGAACTGTGTGGACACCAATGAGTGCCTCAGCCTTGCAGGAACCTGCCTACCCGGCACTTGCCAGAACCTCGAGGGCTC CTTCCGCTGCATCTGTCCCCCTGGCTTCCAGGTGCAGAGTGACCACTGCATTGATATCGACGAGTGCTCAGAGGAGCCCAA CCTCTGCCTCTTTGGCACCTGTACCAACAGCCCTGGGAGCTTCCAGTGCCTCTGCCCACCTGGCTTTGTCCTCTCTGACAA TGGGCACCGTTGCTTTGACACACGGCAGAGTTTCTGCTTCACCCGTTTTGAGGCTGGGAAGTGCTCGGTGCCCAAAGCTTT CAACACCACCAAGACCCGCTGCTGCTGCAGTAAGAGGCCTGGGGAGGGCTGGGGAGACCCCTGCGAACTGTGTCCCCAGGA AGGCAGCGCTGCCTTTCAGGAGCTCTGCCCCTTTGGCCACGGGGCAGTCCCAGGCCCGGATGACTCCCGAGAAGACGTGAA TGAGTGTGCAGAGAACCCTGGCGTCTGCACTAACGGCGTCTGTGTCAACACCGATGGATCCTTCCGCTGTGAGTGTCCCTT TGGCTACAGCCTGGACTTCACTGGCATCAACTGTGAGGACACAGACGAGTGCTCTGTCGGCCACCCCTGTGGGCAAGGGAC ATGCACCAATGTCATCGGAGGCTTCGAATGTGCCTGTGCTGACGGCTTTGAGCCTGGCCTCATGATGACCTGCGAGGACAT CGACGAATGCTCCCTGAACCCGCTGCTCTGTGCCTTCCGCTGCCACAATACCGAGGGCTCCTACCTGTGCACCTGTCCAGC CGGCTACACCCTGCGGGAGGACGGGGCCATGTGTCGAGATGACAATGAATGCCACGCTCAGCCTGACCTCTGTGTCAACGG CCGCTGTGTCAACACCGCGGGCAGCTTCCGGTGCGACTGTGATGAGGGATTCCAGCCCAGCCCCACCCTTACCGAGTGCCG CGACATCCGGCAGGGGCCCTGCTTTGCCGAGGTGCTGCAGACCATGTGCCGGTCTCTGTCCAGCAGCAGTGAGGCTGTCAC CAGGGCCGAGTGCTGCTGTGGGGGTGGCCGGGGCTGGGGGCCCCGCTGCGAGCTCTGTCCCCTGCCCGGCACCTCTGCCTA CAGGAAGCTGTGCCCCCATGGCTCAGGCTACACTGCTGAGGGCCGAGATGTAGATGAATGCCGTATGCTTGCTCACCTGTG TGCTCATGGGGAGTGCATCAACAGCCTTGGCTCCTTCCGCTGCCACTGTCAGGCCGGGTACACACCGGATGCTACTGCTAC TACCTGCCTGGATATGGATGAGTGCAGCCAGGTCCCCAAGCCATGTACCTTCCTCTGCAAAAACACGAAGGGCAGTTTCCT GTGCAGCTGTCCCCGAGGCTACCTGCTGGAGGAGGATGGCAGGACCTGCAAAGACCTGGACGAATGCACCTCCCGGCAGCA CAACTGTCAGTTCCTCTGTGTCAACACTGTGGGCGCCTTCACCTGCCGCTGTCCGCCCGGCTTCACCCAGCACCACCAGGC CTGCTTCGATGTGAATGAATGTGATGGGCCCCACCGCTGCCAGCATGGCTGTCAGAACCAGCTAGGGGGCTACCGCTGCAG CTGCCCCCAGGGTTTCACCCAGCACTCCCAGTGGGCCCAGTGTGTGGATGAGAATGAGTGTGCCCTGTCGCCCCCCACCTG CGGGAGCGCCTCCTGTCGCAACACTCTTGGTGGCTTCCGCTGCGTCTGCCCCTCTGGCTTTGACTTTGATCAGGCCCTCGG GGGCTGCCAGGAGGTGGATGAGTGCGCCGGACGGCGTGGCCCCTGTAGCTACAGCTGTGCCAACACGCCTGGTGGCTTCCT GTGCGGCTGTCCTC_AAGGCTACTTCCGGGCTGGGC_AAGGGCACTGTGTCTCCGGCCTGGGCTTCAGCCCCGGACCCCAGGA CACCCCGGAC_AAAG_AGGAGCTGCTCTCGTCTGAAGCCTGCTACGAATGCAAGATCAATGGCCTCTCCCCTCGGGACCGGCC _ACGACGC_AGTGCCC_ACAGGG_ACC_ACC_AGGTG_AACCTGGCCACCCTTGACTCCGAGGCCCTGCTGACCTTGGGCCTGAACCT CTCACACCTGGGCCGGGCCGAGCGCATCCTGGAGCTCCGGCCGGCCCTGGAGGGTCTAGAGGGCCGGATCCGCTACGTCAT CGTCCGCGGA_AACGAGC_AAGGTTTCTTTCGCATGC_ATCACCTCCGTGGCGTCAGCTCCCTGCAGCTGGGGCGGAGGCGGCC GGGGCCTGG_AACCT_ACCGGCTGG_AGGTGGTG_AGCCACATGGCAGGACCCTGGGGTGTCCAGCAAGAGGGGCAGCCAGGGCC _ATGGGGCC_AGGCCTTGAGGCTGA_AGGTGCAACTGTCAGTTGCTTTAGTTGGGAGGAGCCTCAGTGGGCCCCAGCTGTCCAG _AG_AAGGGGGATTCTGGA_ACTGGG_AAGG_ACTGATCCCCAGAAGCGATGGCTGACCAGATTGAACCCCGAAACTCAGGAAGAG TG_AAATGCT_AC_ACG_ACAACCTC_AGGCAAGCCCGGCCTCTGCCTGGGCCTCTGTGCCAGCCCCGGGGGCCCCCCAGTTACTC _AGTC_TT_TCCTGG_AGACAGC_AAG_AAGCTGC_AATGTGC_AATCCCCCTGCCCCCACAGCCAAGGTCAGGAAGAGGCCCTGTGGT C_ACCGTGTCTGGCC_AATCTCAGGCTTTC_ACTTCTGTACTGCACTGTGGCTTGCCCTGGCGGGGGGCAGGGGGTTGGCAGGA C_ATGGC_AATGGGCAACTGGGGTGGGC_AC_AGGG_CTT_ATTCCTCGGAGTAGAAGGGTGTACAGGGGGCCCAGACTCCACAGTG _ACTTGCC_AC_ATTTGCCCCCCATTTGG_AG_AATGCTTTT_ATATCAAAAGTGGAGACGATAATAAAGTTATTTTGGGTTAAGTC TGCCTGCCCT_TTGGC_AAGTTCTTG_AAGT_AAGT_AG_ATGCTGC_CCTCGGACTGGGCGAGGCAGATCTTGTGCCTGGGGAAGCA GA_AGG_CCTT_ATGGGCTCCCC_AG_AATGG_TAAT_AATGG_CTCACGCTTCCTGACCACGTACTACATACCAGACACCATTCGATT TTTTTTTTATTTTTT_CTG_AG_AC_AGGGTCTTGCTCTGTTGCCTAGGTTGGAGTGCAGTGGCGCGATCGTGGCTCCCTGTGGT CGCC_ACTTTCCGGGCTCG_AGC_AATCCTC_CCACCTCAGCCTCTCCCAAGTAGCTGGGACCAAAGGTGCACGCCACCACACCC _ΆGCTAΆTTTTTTT_AAATTTCTTT_TTCTTTCTCTTTCTTTTTGAGACCAGCCCGGCCAACATGACGAAACCCCATCTCTAΆT

A

The disclosed NO V2f nucleic acid sequence, localized to chromsome 19, has 3401 of 3689 bases (92%) identical to a Homo sapiens KIAAl 776 protein (fibrillin3) mRNA (gb:GENBANK-ID:AB053450|acc:AB053450.2) (E = 0.0).

A NO V2f polypeptide (SEQ ID NO:16) encoded by SEQ ID NO:15 has 2757 amino acid residues and is presented using the one-letter code in Table 2L. Signal P, Psort and/or Hydropathy results predict that NO V2f contains a signal peptide and is likely to be localized to the nucleus with a certainty of 0.6000. Although PSORT suggests that the Fibrillin-like protein may be localized in the nucleus, the NO V2f protein is similar to the Fibrillin family, some members of which are released extracellularly. Therefore it is likely that NO V2f protein shows a similar localization. The most likely cleavage site for a NOV2f peptide is between amino acids 29 and 30, at: AGG-QG.

Table 2L. Encoded NOV2f protein sequence (SEQ ID NO:16).

MTLEGLYLARGPLARLL AWSALLCMAGGQGR DGALEAAGPGRVRRRGSPGILQGPNVCGSRFHAYCCPG RTFPGRSQC VVPICRRACGEGFCSQPNLCTCADGTLAPΞCGVSRAICDRGCHNGGRCIGPNRCACVYGFMGPQCERDYRTGPCFGQVGPE GCQHQLTGLVCTKALCCATVGRAWGLPCELCPAQPHPCRRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCP VGHR SDSSAACEDYDECSTIPGICEGGECTNTVSSYFCKCPPGFYTSPDGTLHGQSRAGACFSV FGGRCAGDLAGHYTR RQCCCDRGRCWAAGPVPELCPPRGSNEFQQ CAQRLPL PGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARGIPSLGP GNSNIGTATLNQTIDICRHFTNLCLNGRCLPTPSSYRCECNVGYTQDVRGECIDVDECTSSPCHHGDCVNIPGTYHCRCYP GFQATPTRQACVDVDECIVSGGLCHLGRCVNTEGSFQCVCNAGFELSPDGKNCVDHNECATSTMCVNGVCLNEDGSFSCLC KPGF LAPGGHYCMDIDECQTPGICVNGHCTNTEGSFRCQC GGLAVGTDGRVCVDTHVRSTCYGAIEKGSCARPFPGTVT KSECCCANPDHGFGEPCQLCPAKDSAEFQA CSSGLGITTDGRDINECALDPEVCANGVCEN RGSYRCVCN GYEAGASG KDCTDVDECALNSL CDNGWCQNSPGSYSCSCPPGFHFWQDTEICKDVDEC ΞSPCVSGVCRNLAGSYTCKCGPGSR DPS GTFCLDSTKGTCW KIQESRCEVNLQGAS RSECCATLGAAWGSPCERCEIDPACARGFARMTGVTCDDVNECESFPGVCP NGRCVNTAGSFRCECPEGLMLDASGRLCVDVRLEPCF R DEDECGVT PGKYRMDVCCCSIGAVWGVECEACPDPESLEF ASLCPRGLGFASRDF SGRPFYKDVNECKVFPGLCTHGTCRNTVGSFHCACAGGFA DAQERNCTDIDECRISPDLCGQGT CVNTPGSFECECFPGYESGFMLMKNCMDVDECARDPLLCRGGTCTNTDGSYKCQCPPGHE TAKGTACEDIDECSLSDG C PHGQCVNVIGAFQCSCHAGFQSTPDRGATSASCPTEGHVQVVLGPGEQMCTGWSIRAKLSTVPNPAPPDVDECEENPRVCD QGHCTNMPGGHRCLCYDGFMATPDMRTCVDVDECDLNPHIC HGDCENTKGSFVCHCQ GYMVRKGATGCSDVDECEVGGH NCDSHASCLNIPGSFSCRCLPGWVGDGFECHDLDECVSQEHRCSPRGDCLNVPGSYRCTCRQGFAGDGFFCEDRDECAENV DLCDNGQCLNAPGGYRCECEMGFDPTEDHRACQDVDECAQENLCAFGSCENLPGMFRCICNGGYE DRGGGNCTDINECAD PVNCINGVCINTPGSYLCSCPQDFELNPSGVGCVDTRAGNCFLETHDRGDSGISCSAEIGVGVTRASCCCS GRAWGNPCE CPMANTTEYRT CPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCECPPGYH SEHTRICEDIDECSTHS GICGPGTCYNTLGNYTCVCPAEYLQVNGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCEACPTP ISPDYQI CGNQAPGF TDIHTGKPLDIDECGEIPAICANGICINQIGΞFRCECPAGFNYNSIL ACEDVDECGSRESPCQ QNADCINIPGSYRCKCTRGYKLSPGGACVGRNECREIPNVCSHGDCMDTEGSYMCLCHRGFQASADQTLCMDIDECDRQPC GNGTCKNIIGSYNC CFPGFVVTHNGDCVDFDECTTLVGQVCRFGHC NTAGSFHCLCQDGFELTADGKNCVDTNECLSLA GTCLPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSPGSFQCLCPPGFVLSDNGHRCFDTRQSFCF TRFEAGKCSVPKAFNTTKTRCCCSKRPGEG GDPCELCPQEGSAAFQELCPFGHGAVPGPPDSREDVNECAENPGVCTNGV CVNTDGSFRCECPFGYSLDFTGINCEDTDECSVGHPCGQGTCTNVIGGFECACADGFEPG MMTCEDIDECSLNP LCAFR CHNTEGSY CTCPAGYT REDGAMCRDDNECHAQPDLCVNGRCVNTAGSFRCDCDEGFQPSPTLTECRDIRQGPCFAEVLQ TMCRS SSΞSEAVTRAECCCGGGRGWGPRCELCP PGTSAYRKLCPHGSGYTAEGRDVDECRMLAH CAHGECINSLGSFR CHCQAGYTPDATATTCLDMDECSQVPKPCTFLCKNTKGSFLCSCPRGYL EEDGRTCKDLDECTSRQHNCQF CVNTVGAF TCRCPPGFTQHHQACFDVNECDGPHRCQHGCQNQLGGYRCSCPQGFTQHSQWAQCVDENECALΞPPTCGSASCRNTLGGFR CVCPSGFDFDQALGGC_QEVDECAGRRGPCSYSCANTPGGFLCGCPQGYFRAGQGHCVSGLGFSPGPQDTPDKEEL SSEAC YECKINGLSPRDRPRRSAHRDHQVNLATLDSEAL T GLNLSHLGRAERILE RPALEGLEGRIRYVIVRGNEQGFFRMHH LRGVSSLQLGRRRPGPGTYRLEVVSHMAGPWGVQQEGQPGPWGQALR KVQLSVALVGRSLSGPQLSREGGFWN EG IPR SDG

The NO V2f amino acid sequence has 2176 of 2568 amino acid residues (84%) identical to, and 2245 of 2568 amino acid residues (87%) similar to, a Homo sapiens 2809 amino acid residue fibrillin 3 protein (ptnr:TREMBLNEW-ACC:BAB47408) (E = 0.0).

NOV2f is expressed in at least the following tissues: Mammalian Tissue, Brain and Lung. Expression information was derived from the tissue sources ofthe sequences that were included in the derivation ofthe NO V2f sequence.

In addition, NOV2f is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Homo sapiens mRNA homolog for KIAAl 776 protein (fibrillin3) (gb:GENBANK-ID:AB053450|acc:AB053450.2): adrenal gland, bone marrow, brain - amygdala, brain - cerebellum, brain - hippocampus, brain - substantia nigra, brain - thalamus, brain -whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma - Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea and uterus.

Possible small nucleotide polymorphisms (SNPs) found for NOV2a are listed in Table 2M.

NOV2a - NOV2f are very closely homologous as is shown in the amino acid alignment in Table 2N.

Table 2N Amino Acid Alignment of NOV2a - NOV2f

10 20 30 40 50 ....|....|....|....|....|....|....|....|....|....| NOV2a MSKSC T LCP QRLAPPGPTWPS PS P PTPVLPWLCTLSPKT LXH

_N0V2b

_NOV2c

NOV2d

NOV2e

_NOV2f

60 70 80 90 100 ....|....|....|....|....|....|....|....|....|....| O 02/066643

NOV2a HTSSRKPPLTVHRFPVGAGCLLGLPEAPGLPHLATTLPSSLCWVPHGXGG NOV2b NOV2C NOV2d NOV2e NOV2f

160 170 180 190 200

310 320 330 340 350

360 370 380 390 400 ..I

NOV2a RRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVGHRLSE NOV2b RRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVGHRLST NOV2c RRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVGHRLSΓ NOV2d RRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPVG"^NΤC" NOV2e RRGFIPNIHTGACQDVDECQAVPGLCQGGSCVNMVGSFHCRCPV^^' NOV2f RRGFI PNIHTGACODVDECOAVPGLCOGGSCVNN

410 420 430 440 450

I

NOV2a fa»^ma tt »tιa» aiM^ι^'MM«MaattxΛβΛa«t»miimw NOV2b NOV2c SSAACEDYDECSTIPGICEGGECTNTVSSYFCKCPPGFYTSPDGTLHGQS NOV2d SSAACEDYDECSTIPGICEGGECTNTVSSYFCKCPPGFYTSPDGTLHGOS NOV2e SSAACEDY^" NOV2f ι3^H8E3B^ΞS3iE^^SSS3BMtB3^8iiBS

460 470 480 490 500

I .. I .. I .. I .. I . .. I .. I ..I

NOV2a RAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSN NOV2b RAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSN NOV2c RAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSN NOV2d RAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSN NOV2e RAGACFSVLFGGRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSN NOV2f RAGACFSVLFGGRCAGDLAGHYTRROCCCDRGRCWAAGPVPELCPPRGSN 510 520 530 540 550

. . I . . I . . | . . I . . | . . . . I . . I . . . I

NOV2a EFQQLCAQRLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARGI NOV2b EFQQLCAQRLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARG3 NOV2C EFQQLCAQRLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARG1 NOV2d ;EFQQLCAQRLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARGJ NOV2e EFQQLCAQRLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARGI NOV2f lEFOOLCAORLPLLPGHPGLFPGLLGFGSNGMGPPLGPARLNPHGSDARGI

760 770 780 790 800

810 820 830 840 850

I . . I . . I I . . I . . . . I . . . . I

NOV2a ECCCANPDHGFGEPCQLCPAKDSAEFQALCSSGLGITTDGRDINECALDP NOV2b ECCCANPDHGFGEPCQLCPAK|SAEFQALCSSGLGITTDGRDINECALDP NOV2c ECCCANPDHGFGEPCQLCPAKDSAEFQALCSSGLGITTDGRDINECALD] NOV2d ECCCANPDHGFGEPCQLCPAKDSAEFQALCSSGLGITTDGRDINECALD1 NOV2e ECCCANPDHGFGEPCQLCPAK^SAEFQALCSSGLGITTDGRDINECALDP NOV2f ECCCANPDHGFGEPCOLCPAKDSAEFOALCSSGLGITTDGRDINECALDi

860 870 880 890 900

I I. ..I.. I . .. I .. I .. I .. I

NOV2a EVCANGVCENLRGSYRCVCNLGYEAGASGKDCTDVDECALNSLLCDNGWC NOV2b EVCANGVCENLRGSYRCVCNLGYEAGASGKDCTDVDECALNSLLCDNGWC NOV2c EVCANGVCENLRGΞYRCVCNLGYEAGASGKDCTDVDECALNSLLCDNGWC NOV2d EVCANGVCENLRGSYRCVCNLGYEAGAΞGKDCTDVDECALNSLLCDNGWC NOV2e EVCANGVCENLRGSYRCVCNLGYEAGASGKDCTDVDECALNSLLCDNG C NOV2f EVCANGVCENTjRGSYRCVCNLGYEAGASGKDCTDVDECALNST.LCDNGWC

910 920 930 940 950

I .. I .. .. I .. I .. I

NOV2a QNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGSYTCK NOV2b QNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGSYTCK NOV2C QNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGSYTCK NOV2d QNSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGSYTCK NOV2e ONSPGSYSCSCPPGFHFWQDTEICKDVDECLSSPCVSGVCRNLAGSYTCK NOV2f

1110 1120 1130 1140 1150

1310 1320 1330 1340 1350

1360 1370 1380 1390 1400 | .... I .... I • • I I . .. | I .. I ...

NOV2a \PPDVDECEENPRVCDQGHCTNMPGGHRCLCYDGF ATPDMRTCVDV1 NOV2b ^DVDECEENPRVCDQGHCTNMPGGHRCLCYDGFMATPDMRTCVDVI NOV2c APPDVDECEENPRVCDOGHCTNMPGGHRCT.CYDGFMATPDMRTCVDVI

1510 1520 1530 1540 1550

1710 1720 1730 1740 1750

NOV2a TEYRTLCPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCE NOV2b TEYRTLCPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCE NOV2c TEYRTLCPGGEGFQPNRITVILEDIDECQELPGLCQGGDCVNTFGSFQCE NOV2d NOV2e NOV2f

1760 1770 1780 1790 1800 .. I ... .. I ... ..|...

NOV2a ;PPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTLGNYTCVC NOV2b :PPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTL< NOV2c JPPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTLGNΪT<_VL NOV2d DPPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTLGNYTCVC NOV2e ;PPGYHLSEHTRICEDIDECSTHSGICGPGTCYNTΓ^^WV'^ΓΓ'"^Γ NOV2f :PPGYHLSEHTRICEDIDECSTHSGICGPGTCYNT]

1810 1820 1830 1840 1850

NOV2a NOV2b NGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRK CCCSYNIGQA NRPCE NOV2c GGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCE NOV2d JGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCE NOV2e IMGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCE NOV2f NGGNNCMDMRKSVCFRHYNGTCQNELAFNVTRKMCCCSYNIGQAWNRPCE

1860 1870 1880 1890 1900

1960 1970 1980 1990 2000

2010 2020 2040 2050

I I ..I... ..I

NOV2a NOV2b ADQTLCMDI— NOV2C ADQTLCMDIDECDRQPCGNG NOV2d ADQTLCMDIDECDRQPCGNGTC NllGSYNe NOV2e ADQTLCMDIDECDRQPCGNGTCKNI IGSYNC NOV2f

2060 2070 2080 2090 2100

2110 2120 2130 2140 2150 I I ... .. I I I ... I .. I .... I .. I

NOV2a CLPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSE NOV2b CLPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSF NOV2c ^LPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSF NOV2d LPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSP NOV2e CLPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSF NOV2f ^"".LPGTCQNLEGSFRCICPPGFQVQSDHCIDIDECSEEPNLCLFGTCTNSF

2160 2170 2180 2190 2200 I I I I I I I I I

NOV2a GSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRC NOV2b GSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRC NOV2C GSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRC NOV2d GSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRC NOV2e GSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRC NOV2f GSFQCLCPPGFVLSDNGHRCFDTRQSFCFTRFEAGKCSVPKAFNTTKTRC

2260 2270 2280 2290 2300 I I I I I I I . . . . I I

N0V2a DVNEC_AE_NPGVCTNGVCVNTDGSFRCECPFGYSLDFTGINCFFIDTDECSVC N0V2b DVNECAENPGVCTNGVCVNTDGSFRCECPFGYSLDFTGINC|DTDECSVG NOV2c DVNECAENPGVCTNGVCVNTDGSFRCECPFGYSLDFTGINC|DTDECSVG NOV2d DVNECAENPGVCTNGVCVNTDGSFRCECPFGYSLDFTGINCBDTDECSVG NOV2e NOV2f

2410 2420 2430 2440 2450

2460 2470 2480 2490 2500

2510 2520 2530 2540 2550

2560 2570 2580 2590 2600

0

2660 2670 2680 2690 2700

₂710 2720 2730 2740 2750 I .... I I .... I ... .. I I I I I

NOV2a GPHRCQHGCQNQLGGYRCSCPQBFTQHSQWAQCVDENECALSPPTCGSAS NOV2b GPHRCQHGCQNQLGGYRCSCPQAFTQHSQWAQCVDENECALSPPTCGSAS NOV2c GPHRCQHGCQNQLGGYRCSCPQAFTQHSQWAQCVDENECALSPPTCGSAS NOV2d GPHRCQHGCQNQLGGYRCSCPQAFTQHSQWAQCVDENECALSPPTCGSAS NOV2e GPHRCQHGCQNQLGGYRCSCPQAFTQHSQWAQCVDENECALSPPTCGSAS NOV2f GPHRCQHGCQNQLGGYRCSCPQffiFTQHSQWAQCVDENECALSPPTCGSAS

2860 2870 2880 2890 2900

2910 2920 2930 2940 2950

Homologies to any ofthe above NOV2 proteins will be shared by the other NOV2 proteins insofar as they are homologous to each other as shown above. Any reference to NOV2 is assumed to refer to both ofthe NOV2 proteins in general, unless otherwise noted.

NOV2a also has homology to the amino acid sequences shown in the BLASTP data listed in Table 2O.

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 2P.

Table 2P. ClustalW Analysis of NOV2a

1) NOV2a (SEQ ID NO:6)

2) gil 1335064lemblCAA45118.11 (X63556) fibrillin [Homo sapiens] (SEQ ID NO:56)

2) gil4557591lref1NP 000129.11 (NM_000138) fibrillin 1; Fibrillin-1 [Homo sapiens] (SEQ ID NO:57)

3) gil663111|gblAAA62177.1 l (U20217) fibrillin-2 [Mus musculus] (SEQ ID NO:58)

4) gil 1706768lsp|P98133IFBN1 BOVIN FIBRILLIN 1 PRECURSOR (MP340) (Bos taurus) (SEQ ID NO:59)

5) gill 3929180lref1NP 114014.11 (NM_031826) fibrillin-2 [Rattus norvegicus] (SEQ ID NO:60)

10 20 30 0 50

.... I I I |.... I I ....I I .... I I

NOV2a MΞKSCMTLLCPLLQRLAPPGPTWPSLPSLPLPTPV PWLC gi|1335064| YFSREKPLRGRYLKRWGKEGAAGAAAETVGATSGQEPQLGQLRAEPSSGC gi I 45575911 gi I 6631111 gi|1706768| gi I 139291801

60 70 80 90 100 I I I I ....I I .... I I I I

NOV2a TLSPKTL XHHTSSRKPPLTVHRFPVGAGCLLGLPEAPGLPHLATTLPSS gi 113350641 SGHD EQPPPPPRESEPPL HWQGPPEVGAAPGEGGRSPARGTGGGIAGP gi I 45575911 gi I 66311 1 g 11706768 I gi 113929180 | -MGR

110 120 130 140 150

gi l 13929180 I Gl^EππFHBpf YfinEWBAB§-- - QQEB

310 320 330 340 350

360 370 380 390 400

410 420 430 440 450

510 520 530 540 550

gi l 663111 1 gi I 1706768 | gi I 13929180 |

860 870 880 890 900

960 970 980 990 1000

1060 1070 1080 1090 1100 I I I I I „•!... • I

NOV2a SΞlSElHvfiE pffl

1360 1370 1380 1390 1400

1460 1470 1480 1490 1500

1510 1520 1530 1540 1550

2210 2220 2230 2240 2250

gi| 17067681 gi I 139291801

2410 2420 2430 2440 2450

2460 2470 2480 2490 2500

2610 2620 2630 2640 2650

gi I 45575911 gi I 6631111 gi I 17067681 |GSA|, gi|13929180| ISSSNl

3010 3020 3030 3040 3050 .1 I I .... I I I I

NOV2a ©fSEGfflPGP GlALPJiKVS JSVF3!VG SLSGPQLSREGGFWN EGLIPRSD gi I 1335064 I RELNiLEDKYDKDYHSGI IGDNHK K'IQVLLH gi I 45575911 SLN@LEDKYDKDYBSG^GDNBKM|IQVLLH gi I 6631111 gi 11706768 | §ELNgLEDKYDKDYfflSGEfflGDNffl |KKMM|g;:IQILLH- gi 113929180 | ggLR LEER EGSYl GVHGEgflRMS I:LQIQLY-

N0V2a gil 1335064 I g I 45575911 gi I 6631111 gil 1706768 I gi 113929180 I

Tables 2Q - 2X list the domain description from DOMAIN analysis results against NOV2a. This indicates that the NOV2a sequence has properties similar to those of other proteins known to contain these domains.

Table 2Q Domain Analysis of NOV2a gnl I Pfam|pfam00683, TB, TB domain. This domain is also known as the 8 cysteine domain. This family includes the hybrid domains. This cysteine rich repeat is found in TGF binding protein and fibrillin.

(SEQ ID NO: 61)

Length = 42 residues, 95.2% aligned

Score = 50.1 bits (118), Expect = 2e-06

NOV2a 1663 CSAEIGVGVTRASCCCSLGRAWGNPCELCPMANTTEYRTL 1702

II + I I ++ I I I I I I I I I I I I I I 1+ I I I + 11 00683 3 CSNPLPGRVTKSECCCSLGRAWGTPCEPCPVPGTAEYKTL 42 Table 2R Domain Analysis of NOV2a qnl|Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 62)

Length = 42 residues, 100.0% aligned

Score = 46.2 bits (108), Expect = 3e-05

NOV2a 2181 G CSVPKAFNTTKTRCCCSKRPGEGWGDPCELCPQEGSAAFQEL 2224 l + l I I I 1+ I I I I I I I I I I I I l + l ++ I 00683 1 GRCSNPLPGRVTKSECCCS—LGRA GTPCEPCPVPGTAEYKTL 42

Table 2S Domain Analysis of NOV2a gnl|Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 63)

Length = 42 residues, 100.0% aligned

Score = 43.9 bits (102) , Expect = le-04

NOV2a 1816 GTCQNELAFNVTRKMCCCSYNIGQA NRPCEACPTPISPDYQIL 1859 l l l l 11+ llll l + l I II I I I I + +1+ I 00683 1 GRCSNPLPGRVTKSECCCSL--GRAWGTPCEPCPVPGTAEYKTL 42

Table 2T Domain Analysis of NOV2a gnl I Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 64)

Length = 42 residues, 95.2% aligned

Score = 43.5 bits (101), Expect = 2e-04

NOV2a 314 CQHQLTGLVCTKALCCATVGRAWGLPCELCPAQPHPCRRGF 354

I + I I I I I + I I ++ 1 I I I I I I I I I + 00683 3 CSNPLPGRV-TKSECCCSLGRA GTPCEPCPVPGTAEYKTL 42

Table 2U Domain Analysis of NOV2a gnl I Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 65) Length = 42 residues, 100.0% aligned Score = 43.1 bits (100), Expect = 2e-04

NOV2a 787 GSCARPFPGTVTKSECCCANPDHGFGEPCQLCPAKDSAEFQAL 829

I l+ l II I I I I I I I 1+ +1 I 1+ I I +M++ I 00683 1 GRCSNPLPGRVTKSECCCSLGR-A GTPCEPCPVPGTAEYKTL 42

Table 2V Domain Analysis of NOV2a gnl I Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 66) Length = 42 residues, 95.2% aligned Score = 42.7 bits (99) , Expect = 3e-04

NOV2a 979 SRCEVNLQGASLRSECCATLGAA GSPCERCEIDPACARG 1018

II I I +11 I I +1 I I I l + l I I I + 00683 1 GRCSNPLPGRVTKSECCCSLGRAWGTPCEPCPVPGTAEYK 40

Table 2W Domain Analysis of NOV2a gnl I Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 67) Length = 42 residues, 100.0% aligned Score = 39.3 bits (90), Expect = 0.003

NOV2a 462 GRCAGDLAGHYTRRQCCCDRGRCWAAGPVPELCPPRGSNEFQQL 505

I 1 + 1+ +1 I I I II | + 00683 1 GRCSNPLPGRVTKSECCCSLGRA --GTPCEPCPVPGTAEYKTL 42

Table 2X Domain Analysis of NOV2a gnl|Pfam|pfam00683, TB, TB domain. (SEQ ID NO: 68)

Length = 42 residues, 97.6% aligned

Score = 38.5 bits (88), Expect = 0.006

NOV2a 1085 ECGVTLPGKYRMDVCCCSIGAVWGVECEACPDPESLEFASL 1125

I 111+ I I I l + l II I I I I I + 1+ +1 00683 2 RCSNPLPGRVTKSECCCSLGRA GTPCEPCPVPGTAEYKTL 42

Fibrillins 1 and 2 are the main constituents ofthe extracellular micro fibrils responsible for the biomechanical properties of most tissues and organs. They are cysteine-rich glycoproteins predominantly made of multiple repeats homologous to the calcium-binding epidermal growth factor module, and are translated as precursor proteins cleaved by furine/PACE-like activities. Fibrillins polymerize extracellularly as parallel bundles of head- to-tail monomers. Binding to calcium rigidifies the structure ofthe monomers and the supramolecular organization ofthe macroaggregates. Elastic fibers form a network that contributes to the elasticity and resilience of tissues such as the skin. Histopatho logic and ultrastructural abnormalities in the elastic fibers have been observed in several diseases ofthe skin and other tissues. Recent cloning of several genes involved in elastic fiber architecture has lead to the approach ofthe study of elastic fiber genodermatoses through molecular analysis. In recent years, mutations in several ofthe genes encoding elastic fiber proteins have been demonstrated in other diseases. Fibrillin-1 mutations result in the pleiotropic manifestations of Marfan syndrome, and fibrillin-2 alterations cause the overlapping phenotype of congenital contractural arachnodactyly, and, most recently, demonstration of abnormalities in the Menkes syndrome gene in X-linked cutis laxa. The first disorders to involve mutations in the elastin gene itself are, surprisingly, cardiovascular and neurobehavioral disorders, such as supravalvular aortic stenosis and Williams syndrome. It is hypothesized that fibrillin-2 guides elastogenesis, whereas fibrillin-1 provides force-bearing structural support (PMID: 10216958, PMID: 7963685).

The above defined information for NOV2 suggests that the NOV2 protein may function as a member of a family of novel Fibrillin-like proteins. Therefore, the NOV2 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV2 compositions ofthe present invention will have efficacy for treatment of patients suffering from connective tissue disorders, such as severe neonatal Marfan syndrome, dominant ectopia lentis, familial ascending aortic aneurysm, isolated skeletal features of Marfan syndrome, and Shprintzen-Goldberg syndrome, genodermatoses, contractural arachnodactyly, inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; blood disorders; asthma; psoriasis; vascular disorders, hypertension, skin disorders, renal disorders including Alport syndrome, immunological disorders, tissue injury, fibrosis disorders, bone diseases, Ehlers- Danlos syndrome type NI, VII, type IV, S-linked cutis laxa and Ehlers-Danlos syndrome type V, osteogenesis imperfecta, Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, inflammation and wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), Parkinson's disease, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, Crohn's disease; multiple sclerosis; and Treatment of Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, benign prostatic hypertrophy, contractural arachnodactyly/CCA, arthrogryposis multiplex congenita, osteogenesis imperfecta, keratoconus, scoliosis, duodenal atresia, esophageal atresia and intestinal malrotation. The NOV2 nucleic acid encoding Fibrillin-like proteins, and the Fibrillin-like proteins ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV3

A disclosed NOV3 nucleic acid of 2713 nucleotides (also referred to as GSAL442663.1 A) encoding a novel KIAA1589-like protein is shown in Table 3A. An open reading frame was identified beginning with a ATG initiation codon at nucleotides 1-3 and ending with a TAG codon at nucleotides 2711-2713. The start and stop codons are in bold letters in Table 3A. Table 3 A. NOV3 Nucleotide Sequence (SEQ ID NO: 17)

ATGGGCCGGGCACGGTGGGTCACATCTGTAATCCCAGCACTTTGGGAGGCCAAGTGTGGGCCTCATTCTAAGTCTGACTT CCCTAATGCTCAGAAGATGGCTATGTTGAGTCCACAAACTTTTGAATTCAGAAACAAGATTGAGTTGATTTCAGAAGCTC TCCCAGAAGATCAAGAAAGAACTTTCCAGGATCTCCAGGAACCTGAATTGAGTCATACGCCAAACAGTGTTCAGAACCCT GTGGAAGTTTCGTGCAGTCTTCAGACTCAAATCTTCGTCTTCACCCCCGGGGCAAGCTCAGTGACTATTATATGGTGGGT GTGTTTCCTTACCAGCGTGAGTATGAGTGCCCAGACTTCCCCAGCAGAGAAGGGCCTGAATCCGGGGCTGATGTGCCAGG AAAGTTACGCTTGCAGCGGGACTGATGAAGCTATCTTTGAGTGTGATGAGTGCTGCAGTCTGCAGTGTCTCCGCTGCGAG GAGGAGCTCCATCGGCAGGAGCGCCTGAGAAACCATGAGCGGATAAGACTCAAACCTGGCCATGTCCCTTACTGTGACCT CTGCAAGGGTCTCAGTGGGCATTTACCAGGTGTTAGGCAGAGGGCAATAGTGAGGTGCCAGACCTGCAAAATTAACTTGT GCCTGGAGTGCCAGAAGAGGACTCATTCTGGGGGTAACAAAAGGAGACACCCTGTTACTGTGTACAATGTCAGTAATCTC CAGGAGTCACTGGAGGCAGAAGAGATGGATGAGGAGACCAAGAGGAAGAAGATGACTGAGAAGGTGGTGAGTTTCCTCCT AGTAGACGAAAATGAAGAAATTCAGGTAACAAATGAAGAAGACTTCATTAGAAAATTGGACTGCAAACCTGATCAGCATC TGAAAGTGGTTTCCATTTTTGGAAATACTGGTGATGGAAAGTCTCATACTCTCAACCACACTTTCTTTTATGGTCGTGAA GTCTTTAAAACCTCCCCGACCCAGGAGTCCTGCACTGTGGGAGTGTGGGCAGCCTATGACCCAGTTCACAAAGTAGCAGT GATCGATACGGAAGGGCTCCTGGGGGCCACCGTGAATCTAAGCCAGAGAACACGGCTGCTGCTTAAGGTCCTGGCCATCT CAGACCTCGTCATCTATCGAACTCATGCAGACCGGCTGCATAACGACCTCTTCAAATTCCTTGGGGATGCCTCAGAAGCT TATCTGAAGCACTTCACCAAGGAGCTCAAGGCCACCACTGCTCGCTGTGGCCTGGATGTCCCTTTATCCACACTGGGCCC TGCAGTTATCATCTTCCATGAGACCGTGCACACCCAGCTACTGGGCTCTGATCATCCCTCAGAGGTGCCAGAGAAGCTCA TCCAGGACCGGTTCCGGAAGCTGGGCCGTTTCCCTGAAGCCTTTAGTTCCATTCACTACAAGGGAACGAGGACTTACAAC CCTCCCACGGACTTTTCTGGGCTTCGGCGTGCTTTGGAACAGCTACTAGAGAATAACACCACCCGTTCTCCCCGGCACCC GGGAGTCATCTTCAAAGCCCTGAAGGCACTAAGTGACCGCTTCAGCGGTGAGATCCCCGATGACCAGATGGCGCACAGCT CCTTTTTTCCAGATGAGTATTTCACCTGCTCCTCCTTGTGCCTCAGCTGTGGGGTTGGATGTAAGAAAAGCATGAATCAT GGGAAGGAAGGAGTGCCTCATGAAGCCAAGAGCCGCTGCAGATACTCCCACCAGTATGACAACCGAGTGTATACCTGCAA GGCCTGCTATGAGAGAGGCGAGGAAGTCAGTGTAGTGCCCAAAACATCTGCTTCCACTGACTCCCCCTGGATGGGTCTCG CAAAATATGCCTGGTCTGGGTATGTGATCGAATGTCCTAACTGTGGCGTGGTCTATCGTAGTCGGCAGTACTGGTTTGGA AACCAAGATCCTGTGGATACGGTGGTGCGGACAGAGATTGTGCATGTGTGGCCTGGAACTGATGGGTTTCTGAAGGACAA CAACAATGCTGCCCAGCGCCTGTTGGACGGGATGAACTTCATGGCTCAGTCGGTGTCCGAGCTTAGCCTTGGACCCACCA AGGCTGTGACTTCCTGGCTGACAGACCAGATCGCCCCTGCCTACTGGAGGCCCAACTCCCAGATTCTGAGCTGCAACAAG TGTGCGACGTCCTTTAAAGATAACGACACTAAGCATCACTGCCGAGCCTGTGGGGAGGGCTTCTGTGACAGCTGTTCATC AAAGACTCGGCCAGTGCCTGAGCGGGGCTGGGGCCCTGCGCCAGTGCGGGTCTGTGACAACTGCTACGAAGCCAGGAACG TCCAGTTAGCTGTTACCGAGGCACAAGTGGACGATGAAGGTGGAACGCTCATTGCTCGGAAGGTGGGCGAGGCCGTGCAG AACACTCTGGGAGCCGTGGTGACAGCCATTGACATACCACTAGGTCTGGTAAAGGACGCGGCCAGGCCTGCGTACTGGGT GCCTGACCACGAAATCCTCCACTGCCACAACTGCCGGAAGGAGTTCAGCATCAAGCTCTCCAAGCACCACTGCCGGGCCT GCGGACAGGGCTTCTGTGATGAGTGCTCCCATGACCGCCGGGCTGTTCCTTCTCGTGGCTGGGACCATCCCGTCCGAGTC TGCTTCAACTGCAATAAAAAGCCCGGTGACCTTTAACCCCAGCCCCCTCTCCGAGTCCTTCACAATTCCTTAG

The disclosed NOV3 nucleic acid sequence maps to chromosome 14 and has 2502 of 2518 bases (99%) identical to a Homo sapiens KIAAl 589 protein mRNA (gb:GENBANK- ID:AB046809|acc:AB046809.1) (E = 0.0).

A disclosed NOV3 protein (SEQ ID NO: 18) encoded by SEQ ID NO: 17 has 891 amino acid residues, and is presented using the one-letter code in Table 3B. Signal P, Psort and/or Hydropathy results predict that NOV3 contains a signal peptide, and is likely to be localized to the nucleus with a certainty of 0.6000 and to the mitochondrial matrix space with a certainty of 0.4811. The most likely cleavage site for a NOV3 peptide is between amino acids 19and 20, at: AKC-GP.

Table 3B. Encoded NOV3 protein sequence (SEQ ID NO:18).

MGRARWVTSVIPALWEAKCGPHSKSDFPNAQKMAMLSPQTFEFRNKIELISEALPEDQERTFQDLQEPELSHTPNSVQNP VEVSCSLQTQIFVFTPGASSVTII VCFLTSVSMSAQTSPAEKGLNPGL CQESYACSGTDEAIFECDECCSLQCLRCE EELHRQERLRNHERIRLKPGHVPYCDLCKGLSGHLPGVRQRAIVRCQTCKINLCLECQKRTHSGGNKRRHPVTVYNVSNL QEΞLEAEEMDEETKRKKMTEKVVSFLLVDENEEIQVTNEEDFIRKLDCKPDQHLKVVSIFGNTGDGKSHTLNHTFFYGRE VFKTSPTQESCTVGV AAYDPVHKVAVIDTEGLLGATVNLSQRTRLLLKVLAISDLVIYRTHADRLHNDLFKFLGDASEA YLKHFTKELKATTARCGLDVPLSTLGPAVIIFHETVHTQLLGSDHPSEVPEKLIQDRFRKLGRFPEAFSSIHYKGTRTYN PPTDFSGLRRALEQLLENNTTRSPRHPGVIFKALKALSDRFSGEIPDDQMAHSSFFPDEYFTCSSLCLSCGVGCKKSMNH GKEGVPHEAKSRCRYSHQYDNRVYTCKACYERGEEVSVVPKTSASTDSPWMGLAKYA SGYVIECPNCGVVYRSRQYWFG NQDPVDTVVRTEIVHVWPGTDGFLKDNNNAAQRLLDGMNFMAQSVSELSLGPTKAVTS LTDQIAPAYWRPNSQILSCNK CATSFKDNDTKHHCRACGEGFCDSCSSKTRPVPERGWGPAPVRVCDNCYEARNVQLAVTEAQVDDEGGTLIARKVGEAVQ NTLGAVVTAIDIPLGLVKDAARPAYWVPDHEILHCHNCRKEFSIKLSKHHCRACGQGFCDECSHDRRAVPSRGWDHPVRV CFNCNKKPGDL The NOV3 amino acid sequence has 816 of 816 amino acid residues (100%) identical to, and 816 of 816 amino acid residues (100%) similar to, a Homo sapiens 816 amino acid residue KIAAl 589 protein (ptnr:TREMBLNEW-ACC:BAB13415) (E = 0.0).

NOV3 is expressed in at least the following tissues: Adrenal Gland/Suprarenal gland, Ascending Colon, Bone, Bone Marrow, Brain, Colon, Heart, Kidney, Liver, Lung, Lymphoid tissue, Mammary gland/Breast, Oesophagus, Ovary, Pituitary Gland, Prostate, Retina, Skeletal Muscle, Small Intestine, Testis, Thyroid, Tongue, Umbilical Vein, Uterus and Whole Organism. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Genomic Clone sources, Literature sources, and/or RACE sources. In addition, NOV3 is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Homo sapiens KIAAl 589 protein mRNA homolog, (GENBANK-ID: gb:GENBANK-ID:AB046809|acc:AB046809.1):Adrenal Gland/Suprarenal gland, Ascending Colon, Bone, Bone Marrow, Brain, Colon, Heart, Kidney, Liver, Lung, Lymphoid tissue, Mammary gland/Breast, esophagus, Ovary, Pituitary Gland, Prostate, Retina, Skeletal Muscle, Small Intestine, Testis, Thyroid, Tongue, Umbilical Vein, Uterus and Whole Organism.

NOV3 also has homology to the amino acid sequences shown in the BLASTP data listed in Table 3C.

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 3D.

Table 3D. ClustalW Analysis of NOV3

1) NOV3 (SEQ ID NO: 18)

2) gι|14749609|ref|XP_027303.1| (XM_027303) hypothetical protein XP_027303 [Homo sapiens] (SEQ ID NO:69)

2) gi|14042048|dbilB AB55085.il (AK027399) unnamed protein product [Homo sapiens] (SEQ ID NO:70)

3) gill0047253|dbilB AB13415.il (AB046809) KIAA1589 protein [Homo sapiens] (SEQ ID NO:71)

4) gι| 14749607|ref1XP_027304.11 (XM_027304) zinc finger protein, subfamily 2A (FYVE domain containing) , 1 [Homo sapiens] (SEQ ID NO:72)

5) gJ|13958036|gb|AAK50771.1|AF361055 1 (AF361055) FYVE-finger protein EIPl [Homo sapiens] (SEQ ID NO:73)

10 20 30 40 50 ....|....|....|....|....|....|....|....| ....I....I

N0V3 GRARWVTSVIPALWEAKCGPHSKSDFPNAQK AMLSPQTFEFRNKIELI

60 70 80 90 100 ....|....|....|....|....|....|....|....|....|....|

NOV3 SEALPEDQERTFQDLQEPELSHTPNSVQNPVEVSCSLQTQIFVFTPGASS gι|147 9609| gι|14042048| gι|10047253| -SVQNPVEVSCSLQTQIFVFTPGASS gi 114749607 | gι|13958036|

160 170 180 190 200

210 220 230 240 250

I ..| .. I .. I .. I ..I .. I. I ..I

NOV3 RAIVRCQTCKINLCLECQKRTHSGGNKRRHPVTVYNVSNLQESLEAEEMD gι | 14749609 | RAIVRCQTCKINLCLECQKRTHSGGNKRRHPVTVYNVSNLQESLEA gi I 14042048 I RAIVRCQTCKINLCLECQKRTHSGGNKRRHPVTVYNVSNLQESI^~ gι|10047253| RAIVRCOTCKINLCLECOKRTHSGGNKRRHPVTVYNVSNLQESLEAEEMD gι|14749607| gι|13958036|

gi|14042048| gi|10047253| gi 114749607 | gi I 13958036 | @RlL|gADHAP[jQQF| v MEHCL|3HGLK gKSF|@QNK|FlGl3lELBEKlC

410 420 430 440 450

I ..|.. .. I .. I .. I .. I .. I .. I ..I

NOV3 SEAYLKHFTKELKATTARCGLDVPLSTLGPAVIIFHETVHTQLLGSDHPS gι|14749609| SEAYLKHFTKELKATTARCGLDVPLSTLGPAVIIFHETVHTQLLGSDHPS gi 114042048 | SEAYLKHFTKELKATTARCGLDVPLSTLGPAVIIFHETVHTQLLGSDHPS gi I 10047253 | SEAYLKHFTKELKATTARCGLDVPLSTLGPAVIIFHETVHTQLLGSDHPS gι|14749607| gι|13958036| JEPEAL EEEGMVIV0jLHG|J vjDAN|clKG|3DlD|)g|GVl|3FSL

460 470 480 490 500 .. I .. I .. I .. I ..I

NOV3 EVPEKLIQDRFRKLGRFPEAFSSIHYKGTRTYNPPTDFSGLRRALEQLL gi I 1474960 | EVPEKLIQDRFRKLGRFPEAFSSIHYKGTRTYNPPTDFSGLRRALEQLLl gi|14042048| EVPEKLIQDRFRKLGRFPEAFSSIHYKGTRTYNPPTDFSGLRRALEQLLI gi 110047253 | EVPEKLIODRFRKLGRFPEAFSSIHYKGTRTYNPPTDFSGLRRALEOLL gi| 14749607 I gi I 139580361 Y|KivQDLBGG^HE|giTlVLDQK|flVEEL RHLSCTVGDi!iTKi G||El

510 520 530 540 550

NOV3 JTTRSPRHPGVIFKALK gi 1147496091 JTTRSPRHPGVIFKALK gi 114042048 | gι|10047253| 'RSPRHPGVIFKALKALSDRFSC DDQMAHSSFFPDEYFTCSSLC gi I 14749607 | MAHSSFFPDEYFTCSSLC gι|13958036| T|J]SKLQEELS- -AATDRICSLQJEgQQQLREQN

560 570 580 590 600

I I .. I .. I I .. I .. I • - I

NOV3 LSCGVGCKKSMNHGKEGVPHEAKSRCRYSHQYDNRVYTCKACYERGEEVS gi 1147496091 LSCGVGCKKSMNHGKEGVPHEAKSRCRYSHQYDNRVYTCKACYERGEEVS gi 114042048 I LSCGVGCKKSMNHGKEGVPHEAKSRCRYSHQYDNRVYTCKACYERGEEVS gi| 100472531 LSCGVGCKKSMNHGKEGVPHEAKSRCRYSHQYDNRVYTCKACYERGEEVS gi|14749607| LSCGVGCKKSMNHGKEGVPHEAKSRCRYSHQYDNRVYTCKACYERGEEVS g 1139580361 ELIRERSEjK^SgEIT|ggDTKVj LE^YKlSTRQGLgE YSDV KQLgEEggHR

610 620 630 640 650

I .. I .. , . I .. I .. I .. I .. I ..

NOV3 VVPKTSASTDSPW GLAKYAWSGYVIECPNCGVVYRSRQYWFGNQDPVDT gi 114749609 | VVPKTSASTDSP MGLAKYAWSGYVIECPNCGVVYRSRQYWFGNQDPVDT gi I 14042048 | VVPKTSASTDSPWMGLAKYAWSGYVIECPNCGVVYRSRQYWFGNQDPVDT gi|10047253| VVPKTSASTDSPWMGLAKYAWSGYVIECPNCGVVYRSRQYWFGNQDPVDT g I 1474960 | VVPKTSASTDSPWMGLAKYAWSGYVIECPNCGVVYRSRQYWFGNQDPVDT gi|13958036| |ELBKELELQIGMKTEMgI>2MKLLEKfTHEK <aDTLVALRQgLEEVK

660 670 680 690 700

I . . I . . I . . I . . I

NOV3 VVRTEIVHVWPGTDGFLKDNNNAAQRLLDG NFMAQSVSELSLGPTKAVT g ) 147496091 WRTEIVHVWPGTDGFLKDNNNAAQRLLDGMNFMAQSVSELSLGPTKAVT gil 14042048 | i/VRTEIVHVWPGTDGFLKDNNNAAQRLLDGMNFMAQSVSELSLGPTKAVT gi|10047253| VVRTEIVHVWPGTDGFLKDNNNAAQRLLDGMNFMAQSVSELSLGPTKAVT gi I 14749607 | J7VRTEIVHVWPGTDGFLKDNNNAAQRLLDGMNFMAQSVSELSLGPTK; gi I 139580361 AlNL^< Eij!κAQNASSSi QK|!]E>3lTSFEGK |5[θ MS@|K^EE LQHlER

710 720 730 740 750

I I . . I . . I . . I . . • • I

NOV3 SWLTDQIAPAYWRPNSQILSCNKCATSFKDNDTKHHCRACGEGFCDSCSS gi| 147496091 SWLTDQIAPAYWRPNSQILSCNKCATSFKDNDTKHHCRACGEGFCDSCSS gi|14042048| SWLTDQIAPAY RPNSQILSCNKCATSFKDNDTKHHCRACGEGFCDSCSS g I 10047253 | S LTDQIAPAYWRPNSQILSCNKCATSFKDNDTKHHCRACGEGFCDSCSS gi 114749607 | SWLTDQIAPAYWRPNSQILSCNKCATSFKDNDTKHHCRACGEGFCDSCSS g I 13958036! -A gG0EE SHKLCφEHGGRIGB^LQ^L§^L§Q^LHEQ!S§ EgELKSEKE) 760 770 780 790 800

I I I ..I • ι . I

810 820 830 840 850

I .. I .. I I ..|.. I .. I .. I .. I

NOV3 AVQNTLGAVVTAIDIPLGLVKDAARPAY :LHCHNCRKEF: gi|14749609| AVQNTLGAVVTAIDIPLGLVKDAARPAY :LHCHNCRKEF: gi|14042048| AVQNTLGAVVTAIDIPLGLVKDAARPAY WVPDHEILHCHNCRKEF: g 110047253 I AVQNTLGAVVTAIDIPLGLVKDAARPAY gi I 14749607 I AVQNTLGAVVTAIDIPLGLVKDAARPAY gi I 139580361 QALQEigLH^QSK|KiED gEVN|ALKGHS

NOV3 g I 147496091 gi| 140420481 gi|10047253| g 114749607 | gil 139580361 TAS

Tables 3E - 3H list the domain description from DOMAIN analysis results against NOV3. This indicates that the NOV3 sequence has properties similar to those of other proteins known to contain these domains.

Table 3E Domain Analysis ofNOV3 gnl | Smart Ismart00064, FYVE, Protein present in Fabl, YOTB, Vacl, and

EEA1; Zinc-binding domain, possibly involved in endo.somal targetting.

Recent data indicates that these domains bind Ptdlns (3)P. (SEQ ID

Nθ:74)

Length = 69 residues, 92.8% aligned

Score = 70.9 bits (172), Expect = 3e-13

NOV3 822 RPAYWVPDHEILHCHNCRKEFSIKLSKHHCRACGQGFCDECSHDRRAVPSRG DHPVRVC

+ 1 + 1 I I + 1 I I I I ++ + I I I I I I + I I + I I + + 1 I + l l l l l

00064 2 VRPHWIPDVEASNCMGCGKEFNLTKRRHHCRNCGRIFCSKCSSKKAPLPKLGNEDPVRVC 61

NOV3 882 F CN 885

+ 1 00064 62 DDCY 65

Table 3F Domain Analysis of NOV3 gnl I Smart I smart00064 , FYVE. (SEQ ID NO: 75) Length = 69 residues, 89.9% aligned Score = 62.8 bits (151), Expect = 8e-ll

NOV3 708 Y RPNSQILSCNKCATSFKDNDTKHHCRACGEGFCDSCSSKTRPVPERG GPAPVRVCDN 767

+ 1 1+ + +1 I I +1 I I I I I II MM 1 + 1+ I I II I 11 + 00064 5 HWIPDVEASNCMGCGKEFNLTKRRHHCRNCGRIFCSKCSSKKAPLPKLGNED-PVRVCDD 63

NOV3 768 CYE 770

I I I 00064 64 CYE 66 Table 3G Domain Analysis of NOV3 gnl I Pfam|pfam01363, FYVE, FYVE zinc finger. The FYVE zinc finger is named after four proteins that it has been found in: Fabl, YOTB/ZK632.12, Vacl, and EEA1. The FYVE finger has been shown to bind two Zn++ ions. The FYVE finger has eight potential zinc coordinating cysteine positions. Many members of this family also include two histidines in a motif R+HHC+XCG, where + represents a charged residue and X any residue. We have included members which do not conserve these histidine residues but are clearly related. (SEQ ID NO: 76) Length = 66 residues, 92.4% aligned Score = 67.4 bits (163), Expect = 3e-12

NOV3 824 AYWVPDHEILHCHNCRKEFSIKLSKHHCRACGQGFCDECSHDRRAVPSRGWDHPVRVCFN 883

+ MM I+ +I I I I++ +1 I I I I I 1+ I I II +1 I + I I I I I + 01363 1 PHWVPDEEVSNCMRCGKPFTLTKRRHHCRACGRIFCSSCSSKTVPLPPMG-ERPVRVCDS 59 NOV3 884 CN 885 I 01363 60 CY 61

Table 3H Domain Analysis of NOV3 gnl I Pfam|pfam01363, FYVE, FYVE zinc finger. (SEQ ID NO: 77) Length = 66 residues, 98.5% aligned Score = 64.7 bits (156), Expect = 2e-ll

NOV3 707 AYWRPNSQILSCN CATSFKDNDTKHHCRACGEGFCDSCSSKTRPVPERGWGPAPVRVCD 766

+ I I+ ++ +I +I I +IIIMM M MMM I + I I I I I I I I 01363 1 PHWVPDEEVSNCMRCGKPFTLTKRRHHCRACGRIFCSSCSSKTVPLPPM--GERPVRVCD 58

NOV3 767 NCYEARN 773

+ 11+ I 01363 59 SCYDLLN 65

The above defined information for NOV3 suggests that this NOV3 protein may function as a member of a KIAAl 589 protein family. Therefore, the NOV3 nucleic acids and proteins ofthe invention are useful in potential therapeutic and diagnostic applications. For example, a cDNA encoding the NOV3 protein may be useful in gene therapy, and the NOV3 protein may be useful when administered to a subject in need thereof. By way of nonlimiting example, the compositions ofthe present invention will have efficacy for treatment of patients suffering from inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; blood disorders; asthma; psoriasis; vascular disorders, hypertension, skin disorders, renal disorders including Alport syndrome, immunological disorders, tissue injury, fibrosis disorders, bone diseases, Ehlers- Danlos syndrome type VI, VII, type IV, S-linked cutis laxa and Ehlers-Danlos syndrome type V, osteogenesis imperfecta, Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, inflammation and wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), Parkinson's disease, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis; multiple sclerosis; and Treatment of Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers and benign prostatic hypertrophy. The NOV3 nucleic acid encoding KIAA1589-like protein, and the KIAAl 589-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV4

A disclosed NOV4 nucleic acid of 1761 nucleotides (designated CuraGen Ace. No. GSAL442663.1_B) encoding a novel WD40-motifprotein-like protein is shown in Table 4A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 48-50and ending with a TAA codon at nucleotides 1536-1538. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 4A, and the start and stop codons are in bold letters.

Table 4A. NOV4 Nucleotide Sequence (SEQ ID NO:19)

AGCTGAGCCGGAGGGAATCCGGAAGGACACGCTGAACAGGΆACAGAAATGAATAAAAGTCGCTGGCAGAGTAGAAGACG ACATGGGAGAAGAAGCCACCAGCAGAACCCTTGGTTCAGACTCCGTGATTCTGAAGACAGGTCTGACTCCCGGGCAGCA CAGCCCGCTCACGATTCCGGCCACGGTGATGACGAGTCTCCGTCAACCTCGTCTGGCACAGCTGGGACCTCCTCTGTGC CAGAGCTACCTGGGTTTTACTTTGACCCTGAAAAGAAACGCTACTTCCGCTTGCTCCCTGGACATAACAACTGCAACCC CCTGACGAAAGAGAGCATCCGGCAGAAGGAGATGGAGAGCAAGAGACTGCGGCTGCTCCAGGAAGAAGACAGACGGAAA AAGATTGCCAGGATGGGATTTAATGCATCTTCCATGCTACGAAAAAGCCAGCTGGGTTTTCTCAACGTCACCAATTACT GCCATTTAGCCCACGAGCTGCGTCTCAGCTGCATGGAGAGGAAAAAGGTCCAGATTCGAAGCATGAGGGATCCCTCCGC CTTGGCAAGCGACCGATTTAACCTCATACTGGCAGATACCAACAGTGACCGGCTCTTCACAGTGAACGATGTTAAAGTT GGAGGCTCCAAGTATGGTATCATCAACCTGCAAAGTCTGAAGACCCCTACGCTCAAGGTGTTCATGCACGAAAACCTCT ACTTCACCAACCGGAAGGTGAATTCGGTGTGCTGGGCCTCGCTGAATCACTTGGATTCCCACATTCTGCTATGCCTCAT GGGACTCGCAGAGACTCCAGGCTGTGCCACCCTGCTCCCAGCATCACTGTTCGTCAATAGTCACCCAGGAATAGACCGG CCTGGCATGCTCTGCAGTTTCCGGATCCCTGGTGCCTGGTCCTGTGCCTGGTCCCTGAATATCCAAGCAAATAACTGCT TCAGTACAGGCTTGTCTCGGCGGGTCCTGTTGACCAACGTGGTGACGGGACACCGGCAGTCCTTTGGGACCAACAGTGA TGTCTTGGCCCAGCAGTTTGCTCTCATGGCTCCTCTGCTGTTTAATGGCTGCCGCTCTGGGGAAATCTTTGCCATTGAT CTGCGTTGTGGAAATCAAGGCAAGGGATGGAAGGCCACCCGCCTGTTTCATGATTCAGCAGTGACCTCTGTGCGGATCC TCCAAGATGAGCAATACCTGATGGCTTCAGACATGGCTGGAAAGATCAAGCTGTGGGACCTGAGGACCACGAAGTGCGT AAGGCAGTACGAAGGCCACGTGAATGAGTACGCCTACCTGCCCCTGCATGTGCACGAGGAAGAAGGAATCCTGGTGGCA GTGGGCCAGGACTGCTACACGAGAATCTGGAGCCTCCACGATGCCCGCCTACTGAGAACCATACCCTCCCCGTACCCTG CCTCCAAGGCCGACATTCCCAGTGTGGCCTTCTCGTCGCGGCTGGGGGGCTCCCGGGGCGCGCCGGGGCTGCTCATGGC TGTCGGGCAGGACCTTTACTGTTACTCCTACAGCTAΛTTCTGCAGGGCACΆGCCCAGAGCCATGTGGATTTGACTTACG GGAGTAAAGCGTAACTTTTTACTGCATCTAATGAGGGTGTTTTAAGTGACACTCAGTGTACACAGATCCCATCCTCTGG CTGCTAGGAGΆGAAGTGCTGAATGTTCCGTGTGGAGATGCTCAGGAAAGTTATTTGAGTTAAATTGCTGGCTGAGAGAG CTTGGAAGTCCTTTTCATAAAAG

The nucleic acid sequence ofNOV4 maps to chromosome 14 and has 1282 of 1287 bases (99%) identical to a Homo sapiens DKFZp434Kl 14 mRNA (gb.GENBANK- ID:HSM800674|acc:AL080157.1) (E = 2.6e-²⁸²). A NOV4 polypeptide (SEQ ID NO:20) encoded by SEQ ID NO: 19 is 496 amino acid residues and is presented using the one letter code in Table 4B. Signal P, Psort and/or Hydropathy results predict that NOV4 does not contain a signal peptide and is likely to be localized to the nucleus with a certainty of 0.9600.

Table 4B. NOV4 protein sequence (SEQ ID NO:20)

MNKSRWQSRRRHGRRSHQQNPWFRLRDSEDRSDSRAAQPAHDSGHGDDESPSTSSGTAGTSSVPELPGFYFDPEKKRYFRLLP GHNNCNPLTKESIRQKEMESKRLRLLQEEDRRKKIARMGFNASSMLRKSQLGFLNVTNYCHLAHELRLSCMERKKVQIRS RD PSALASDRFNLILADTNΞDRLFTVNDVKVGGSKYGI INLQSLKTPTLKVFMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG LAETPGCATLLPASLFVNSHPGIDRPGMLCSFRI PGAWSCAWSLNIQANNCFSTGLSRRVLLTNVVTGHRQSFGTNSDVLAQQ FALMAPLLFNGCRSGEI FAIDLRCGNQGKG KATRLFHDSAVTSVRILQDEQYLMASDMAGKIKL DLRTTKCVRQYEGHVNE YAYLPLHVHEEEGILVAVGQDCYTRI SLHDARLLRTI PSPYPASKAPIPSVΆFSSRLGGSRGAPGLLMAVGQDLYCYSYS

The NOV4 amino acid sequence has 425 of 428 amino acid residues (99%) identical to, and 425 of 428 amino acid residues (99%) similar to, a Homo Sapiens 430 amino acid residue hypothetical 48.5 kDa protein (ptnr:SPTREMBL-ACC:Q9Y4P5) (E = l.Oe^"232).

NOV4 is expressed in at least the following tissues: cancer tissue, pancreas, fetal lung NbHL19W, testis NHT, B-cell and brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Genomic Clone sources, Literature sources, and/or RACE sources. In addition, NOV4 is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Homo sapiens DKFZp434Kl 14 mRNA homolog (GENBANK-ID: gb:GENBANK-ID: HSM800674 |acc:AL080157.1):

Rhabdomyosarcoma, neuroepithelium, pancreas, fetal lung, NbHL19W, testis NHT, B-cell and brain.

NOV4 also has homology to the amino acid sequences shown in the BLASTP data listed in Table 4C.

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 4D.

Table 4D ClustalW Analysis of NOV4

1) NOV4 (SEQ ID NO:20)

2) gi|14749605lref_|XP_027300.1 | (XM_027300) hypothetical protein XP_027300 [Homo sapiens] (SEQ ID NO:78)

3) gi| 14042651 |dbj|BAB55337.11 (AK027745) unnamed protein product [Homo sapiens] (SEQ ID NO:79)

4) gi|7512629|pirllT12541 hypothetical protein DKFZp434Kl 14.1 -(fragment) [Homo sapiens] (SEQ ID NO:80)

5) gill 309698 l |gblAAH03284.1 |AAH03284 (BC003284) Unknown (protein for MG 7874) [Mus musculus] (SEQ ID NO:81)

6) gi|14749602|ref1XP 027301.11 (XM_027301) DKFZP434K114 protein [Homo sapiens] (SEQ ID NO:82)

10 20 30 40 50

110 120 130 140 150

160 170 180 190 200

210 220 230 240 250

I ..I .. I ..|.. ..|.. ..I ..

NOV4 YGIINLQSLKTPTLKVFMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG gi 114749605 | YGIINLQSLKTPTLKVFMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG gi|14042651| YGIINLQSLKTPTLKVFMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG gi 17512629 I YGIINLQSLKTPTLKVFMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG g 1130969811 YGIINLC^LKgPTJ l]vgMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG gi|14749602| YGIINLQSLKTPTLKVFMHENLYFTNRKVNSVCWASLNHLDSHILLCLMG

310 320 330 340 350 I I I I I I I I I I

NOV4 IM^^M^_;t»*fc^HWWn:%M^Jlk<.<i»J«B13aτ.MAl_!l*^i[^,ffl^

61

410 420 430 440 450

510 520 . . I . . . . I .

NOV4 gi I 14749605 | gi|14042651| gi|7512629| gi|13096981| CPDSQEEGRWELPSVSNEDIL g I 14749602 I

Recent studies have demonstrated the importance of F-box/WD40 proteins in the regulation of developmental processes, by a mechanism of specific ubiquitinization and subsequent proteo lysis of target proteins belonging to the Wnt, Hh and NF-kappaB signaling pathways. A new human gene, Dactylin, encoding a novel member ofthe F-box/WD40 protein family was recently cloned and characterized. The Dactylin gene comprises nine exons distributed in more than 85 kb of genomic DNA and encoding a protein with four WD40 repeats and an F-box motif. The chromosomal location of Dactylin and its putative function as an F-box/WD40 repeat protein, likely to be involved in key signaling pathways crucial for normal limb development, make it a promising candidate gene for SHFM3 (lanakiev et al., A novel human gene encoding an F-box/WD40 containing protein maps in the SHFM3 critical region on 10q24. Biochem Biophys Res Commun 261(l):64-70, 1999).

Several other proteins have been identified as containing WD40 repeat domains including the COP1 (constitutively photomorphogenic 1) protein in Arabidopsis which is a repressor of light-regulated development and it mammalian homologue which acts within the nucleus to repress photomorphogenic development (Wang et al., Evidence for functional conservation of a mammalian homologue ofthe light-responsive plant protein COP1. Curr Biol 9(13):711-4, 1999). Another protein which contains the WD40 repeat domain is the Schizosaccharomyces pombe 72 kDa TFIID subunit. This protein contains several significant highly conserved regions including the WD40 repeats, that are indispensable for the viability (Yamamoto et al., Molecular genetic elucidation ofthe tripartite structure ofthe Schizosaccharomyces pombe 72 kDa TFIID subunit which contains a WD40 structural motif. Genes Cells 2(4):245-54, 1997).

The above defined information for NON4 suggests that this ΝOV4 protein may function as a member of a WD40-motif protein-like protein family. Therefore, the NOV4 nucleic acids and proteins ofthe invention are useful in potential therapeutic and diagnostic applications. For example, a cDNA encoding the NOV4 protein may be useful in gene therapy, and the NOV4 protein may be useful when administered to a subject in need thereof. By way of nonlimiting example, the compositions ofthe present invention will have efficacy for treatment of patients suffering from cancer, trauma, regeneration (in vitro and in vivo), viral/bacterial/parasitic infections DiabetesNon Hippel-Lindau (NHL) syndrome , Pancreatitis, Obesity Systemic lupus erythematosus , Autoimmune disease, Asthma, Emphysema, Scleroderma, allergy, ARDS, Von Hippel-Lindau (VHL) syndrome , Alzheimer's disease, Stroke, Tuberous sclerosis, hypercalceimia, Parkinson's disease, Huntington's disease, Cerebral palsy, Epilepsy, Lesch-Νyhan syndrome, Multiple sclerosis, Ataxia-telangiectasia, Leukodystrophies, Behavioral disorders, Addiction, Anxiety, Pain, Neuroprotection Fertility Myasthenia gravis, Leukodystrophies, Pain, Neuroprotection

Endocrine dysfunctions, Diabetes, obesity, Growth and reproductive disorders Hemophilia, Hypercoagulation, Idiopathic thrombocytopenic purpura , Immunodeficiencies and Graft vesus host. The NOV4 nucleic acid encoding WD40-motif protein-like protein, and the WD40-motif protein-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV5

NOV5 includes four novel Opioid Binding Cell Adhesion Molecule-like proteins disclosed below. The disclosed proteins have been named NOV5a, NOV5b, NOV5c and NOV5d.

NOV5a

A disclosed NOV5a nucleic acid of 1018 nucleotides (also referred to as 139785504) encoding a novel Opioid Binding Cell Adhesion Molecule-like protein is shown in Table 5 A. An open reading frame lacking a signal peptide was identified beginning with an CTG at nucleotides 1-3 and ending with a TAG codon at nucleotides 958-960. A putative untranslated region downstream from the termination codon are underlined in Table 5A, and the start and stop codons are in bold letters.

Table 5A. NOV5a Nucleotide Sequence (SEQ ID NO:21)

CTGGCCGGCTTGGCCGTCATCAGCCGAGGGCTGCTCTCCCAGAGCCTGGAGTTCAACTCTCCTGCCGACAACTACACAGTGT GTGAAGGTGACAACGCCACCCTCAGCTGCTTCATCGACGAGCACGTGACCCGCGTGGCTTGGCTGAACCGCTCCAACATCCT GTATGCCGGCAATGACCGCTGGACCAGCGACCCGCGGGTGCGGCTGCTCATCAACACCCCCGAGGAGTTCTCCATCCTCATC ACCGAGGTGGGGCTCGGCGACGAGGGCCTCTACACCTGCTCCTTCCAGACCCGCCACCAGCCGTACACCACTCAGGTCTACC TCATTGTCCACGTCCCTGCCCGCATTGTGAACATCTCGTCGCCTGTGACGGTGAATGAGGGGGGCAATGTGAACCTGCTTTG CCTGGCCGTGGGGCGGCCAGAGCCCACGGTCACCTGGAGACAGCTCCGAGACGGCTTCACCTCGGAGGGAGAGATCCTGGAG ATCTCTGACATCCAGCGGGGCCAGGCCGGGGAGTATGAGTGCGTGACTCACAACGGGGTTAACTCGGCGCCCGACAGCCGCC GCGTGCTGGTCACAGTCAACTATCCTCCGACCATCACGGACGTGACCAGCGCCCGCACCGCGCTGGGCCGGACCGCCCTCCT GCGCTGCGAAGCCATGGCGGTTCCCCCCGCGGATTTCCAGTGGTACAAGGATGACAGACTGCTGAGCAGCGGCACGGCCGAA GGCCTGAAGGTGCAGACGGAGCGCACCCGCTCGATGCTTCTCTTTGCCAACGTGAGCGCCCGGCATTACGGCAACTATACGT GTCGCGCCGCCAATCGACTGGGAGCGTCCAGCGCCTCCATGCGGCTCCTGCGCCCAGGATCCCTGGAGAACTCAGCCCCGAG GCCCCCAGGGCTCCTGGCCCTCCTCTCCGCCCTGGGCTGGCTGTGGTGGAGAATGTAGGCGCAACCCAGTCCAGCTCACCTC CCCCTGCAGGGGGCCTCAAACCAAGAGTGAGAGA

The NOV5a nucleic acid was identified on chromosome 7 and has 260 of 389 bases (66%) identical to a Homo sapiens (clone pHOM) opioid-binding cell adhesion molecule mRNA (gb:GENBANK-ID:HUMOBCAM|acc:L34774.1) (E = 3.4e^*50).

A disclosed NOV5a polypeptide (SEQ ID NO:22) encoded by SEQ ID NO:21 is 319 amino acid residues and is presented using the one-letter code in Table 5B. Signal P, Psort and/or Hydropathy results predict that NOV5a does not contain a signal peptide and is likely to be localized in the mitochondrial matrix space with a certainty of 0.4686 and the cytoplasm with a certainty of 0.4500.

Although SignalP, Psort and/or hydropathy suggest that NOV5a protein may be localized in the mitochondrial matrix space and cytoplasm, the protein predicted here is similar to the Opioid Binding Cell Adhesion Molecule family, some members of which are released extracellularly. The closest homolog SWISSPROT-ACC:Q14982 opioid binding protein/cell adhesion molecule is a type la membrane protein that is localized to the plasma membrane extracellularly. This indicates that the signal peptide ofthe mature protein is cleaved. Therefore it is likely that NOV5a protein is available at the same sub-cellular localization and hence accessible to a diagnostic probe and for various therapeutic applications. It also indicates that the use of a heterologous signal peptide to target the novel protein to the appropriate location, i.e. extracellularly, is appropriate.

Table 5B. Encoded NOV5a protein sequence (SEQ ID NO:22)

LAGLAVISRGLLSQSLEFNSPADNYTVCEGDNATLSCFIDEHVTRVAWLNRSNILYAGNDR TSDPRVRLLINTPEEFSIL ITEVGLGDEGLYTCSFQTRHQPYTTQVYLIVHVPARIVNISSPVTVNEGGNVNLLCLAVGRPEPTVTWRQLRDGFTSEGEI LEISDIQRGQAGEYECVTHNGVNSAPDSRRVLVTVNYPPTITDVTSARTALGRTALLRCEAMAVPPADFQ YKDDRLLSSG TAEGLKVQTERTRSMLLFANVSARHYGNYTCRAANRLGASSASMRLLRPGΞLENSAPRPPGLLALLSALGWLWWRM The NOV5a amino acid sequence has 162 of 300 amino acid residues (54%) identical to, and 213 of 00 amino acid residues (71%) similar to, a Homo Sapiens 345 amino acid residue opioid binding protein/cell adhesion molecule precursor (OBCAM) / opioid-binding cell adhesion molecule (OPCML) protein (ptnr.SWISSPROT-ACC:Q14982) (E = 3.1e-⁸⁰). NOV5a is expressed in at least the following tissues: Brain and Fetal brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, genomic clone sources, literature sources, and/or RACE sources. In addition, NOV5a is predicted to be expressed in brain tissues because ofthe expression pattern of a closely related Human (clone pHOM) opioid-binding cell adhesion molecule mRNA homo log (gb:GENBANK- ID:HUMOBCAM|acc:L34774.1).

NOV5b

A disclosed NOV5b nucleic acid of 1017 nucleotides (also referred to as 139785504_dal) encoding a novel Opioid Binding Cell Adhesion Molecule-like protein is shown in Table 5C. An open reading frame lacking the signal peptide was identified beginning with an GCC at nucleotides 3-5 and ending with a TAG codon at nucleotides 958-960. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 5C, and the start and stop codons are in bold letters.

Table 5C. NOV5b Nucleotide Sequence (SEQ ID NO:23)

CTGGCCGGCTTGGCCGTCATCAGCCGAGGGCTGCTCTCCCAGAGCCTGGAGTTCAACTCTCCTGCCGACAACTACACAG TGTGTGAAGGTGACAACGCCACCCTCAGCTGCTTCATCGACGAGCACGTGACCCGCGTGGCCTGGCTGAACCGCTCCAA CATCCTGTATGCCGGCAATGACCGCTGGACCAGCGACCCGCGGGTGCGGCTGCTCATCAACACCCCCGAGGAGTTCTCC ATCCTCATCACCGAGGTGGGGCTCGGCGACGAGGGCCTCTACACCTGCTCCTTCCAGACCCGCCACCAGCCGTACACCA CTCAGGTCTACCTCATTGTCCACGTCCCTGCCCGCATTGTGAACATCTCGTCGCCTGTGACGGTGAATGAGGGGGGCAA TGTGAACCTGCTTTGCCTGGCCGTGGGGCGGCCAGAGCCCACGGTCACCTGGAGACAGCTCCGAGACGGCTTCACCTCG GAGGGAGAGATCCTGGAGATCTCTGACATCCAGCGGGGCCAGGCCGGGGAGTATGAGTGCGTGACTCACAACGGGGTTA ACTCGGCGCCCGACAGCCGCCGCGTGCTGGTCACAGTCAACTATCCTCCGACCATCACGGACGTGACCAGCGCCCGCAC CGCGCTGGGCCGGGCCGCCCTCCTGCGCTGCGAAGCCATGGCGGTTCCCCCCGCGGATTTCCAGTGGTACAAGGATGAC AGACTGCTGAGCAGCGGCACGGCCGAAGGCCTGAAGGTGCAGACGGAGCGCACCCGCTCGATGCTTCTCTTTGCCAACG TGAGCGCCCGGCATTACGGCAACTATACGTGTCGCGCCGCCAACCGACTGGGAGCGTCCAGCGCCTCCATGCGGCTCCT GCGCCCAGGATCCCTGGAGAACTCAGCCCCGAGGCCCCCAGGGCTCCTGGCCCTCCTCTCCGCCCTGGGCTGGCTGTGG TGGAGAATGTAGGCCAACCCAGTCCAGCTCACCTCCCCCTGCAGGGGGCCTCAAACCAAGAGTGAGAGA

The NOV5b nucleic acid was identified on chromosome 7 and has 261 of 389 bases

(67%) identical to a Homo sapiens (clone pHOM) opioid-binding cell adhesion molecule mRNA (gb:GENBANK-ID:HUMOBCAM|acc:L34774.1) (E = 1.3e^"50).

A disclosed NOV5b polypeptide (SEQ ID NO:24) encoded by SEQ ID NO:23 is 319 amino acid residues and is presented using the one-letter code in Table 5D. Signal P, Psort and/or Hydropathy results predict that NOV5b does not contain a signal peptide and is likely to be localized in the mitochondrial matrix space with a certainty of 0.4686 and the cytoplasm with a certainty of 0.4500. Although SignalP, Psort and/or hydropathy suggest that NOV5b protein may be localized in the mitochondrial matrix space and cytoplasm, the protein predicted here is similar to the Opioid Binding Cell Adhesion Molecule family, some members of which are released extracellularly. The closest homolog SWISSPROT-ACC.-Q14982 opioid binding protein cell adhesion molecule is a type la membrane protein that is localized to the plasma membrane extracellularly. This indicates that the signal peptide ofthe mature protein is cleaved. Therefore it is likely that NOV5b protein is available at the same sub-cellular localization and hence accessible to a diagnostic probe and for various therapeutic applications. It also indicates that the use of a heterologous signal peptide to target the novel protein to the appropriate location, i.e. extracellularly, is appropriate.

Table 5D. Encoded NOV5b protein sequence (SEQ ID NO:24).

LAGLAVISRGLLSQSLEFNSPADNYTVCEGDNATLSCFIDEHVTRVAWLNRSNILYAGNDRWTSDPRVRLLINTPEEFSILITEV GLGDEGLYTCSFQTRHQPYTTQVYLIVHVPARIVNISSPVTVNEGGNVNLLCLAVGRPEPTVTWRQLRDGFTSEGEILEISDIQR GQAGEYECVTHNGVNSAPDSRRVLVTVNYPPTITDVTSARTALGRAALLRCEAMAVPPADFQWYKDDRLLSSGTAEGLKVQTERT RSMLLFANVSARHYGNYTCRAANRLGASSASMRLLRPGSLENSAPRPPGLLALLSALGWLWWRM

The NOV5b amino acid sequence has 164 of 300 amino acid residues (54%) identical to, and 213 of 300 amino acid residues (71%) similar to, a Bos taurus 345 amino acid residue opioid binding protein/cell adhesion molecule precursor (OBCAM) / opioid-binding cell adhesion molecule (OPCML) protein (ptnr:SWISSPROT-ACC:Pl 1834) (E = 1.7e^"80).

NOV5b is expressed in at least the following tissues: Brain and Fetal brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, genomic clone sources, literature sources, and/or RACE sources. In addition, NOV5b is predicted to be expressed in brain tissues because ofthe expression pattern of a closely related Homo sapiens (clone pHOM) opioid-binding cell adhesion molecule mRNA homolog (gb:GENBANK- ID:HUMOBCAM|acc:L34774.1).

NOV5c A disclosed NOV5c nucleic acid of 1136 nucleotides (also referred to as CG51027-03) encoding a novel Opioid Binding Cell Adhesion Molecule-like protein is shown in Table 5E. An open reading frame lacking a signal peptide was identified beginning with an TCC at nucleotides 2-4and ending with a TAG codon at nucleotides 923-925. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 5E, and the start and stop codons are in bold letters. Table 5E. NOV5c Nucleotide Sequence (SEQ ID NO:25)

CTCCCAGAGCCTGGAGTTCAACTCTCCTGCCGACAACTACACAGTGTGTGAAGGTGACAACGCCACCCTCAGCTGCTTCATC GACGAGCACGTGACCCGCGTGGCCTGGCTGAACCGCTCCAACATCCTGTATGCCGGCAATGACCGCTGGACCAGCGACCCGC GGGTGTGGCTGCTCATCAACACCCCCGAGGAGTTCTCCATCCTCATCACCGAGGTGGGGCTCGGCGACGAGGGCCTCTACAC CTGCTCCTTCCAGACCCGCCACCAGCCGTACACCACTCAGGTCTACCTCATTGTCCACGTCCCTGCCCGCATTGTGAACATC TCGTCGCCTGTGATGGTGAATGAGGGGGGCAATGTGAACCTGCTTTGCCTGGCCGTGGGGCGGCCAGAGCCCACGGTCACCT GGAGACAGCTCCGAGACGGCTTCACCTCGGAGGGAGAGATCCTGGAGATCTCTGACATCCAGCGGGGCCAGGCCGGGGAGTA TGAGTGCGTGACTCACAACGGGGTTAACTCGGCGCCCGACAGCCGCCGCGTGCTGGTCACAGTCAACTATCCTCCGACCATC ACGGACGTGACCAGCGCCCGCACCGCGCTGGGCCGGGCCGCCCTCCTGCGCTGCGAAGCCATGGCGGTTCCCCCCGCGGATT TCCAGTGGTATAAGGATGACAGACTGCTGAGCAGCGGCACGGCCGAAGGCCTGAAGGTGCAGATGGAGCGCACCCGCTCGAT GCTTCTCTTTGCCAACGTGAGCGCCCGGCATTACGGCAACTATACGTGTCGCGCCGCCAACCGACTGGGAGCGTCCAGCGCC TCCATGCGGCTCCTGCGCCCAGGATCCCTGGAGAACTCAGCCCCGAGGCCCCCAGGGCTCCTGGCCCTCCTCTCCGCCCTGG GCTGGCTGTGGTGGAGAATGTAGGCGCAACCCAGTGGAGCTCACCTCCCCCTGCAGGGGGCCTCAGGCCAAGAGTGAGAGAA ACGGGGGAGCAAGAGCCGTGGGTCTCGTGGGGGCAGAAGAGCTCTCGGCCACCAAGGAAGAAGAGAGAGGAGAAGAGGAGGA GGCAGAGGAAGAAAGATCTTCAGAGAACCCATCACTGTGAGGGATAACGCAAAATTATGCATCTTTCTAC

The NOV5c nucleic acid was identified on chromosome 7 and has 274 of 389 bases (70%) identical to a Bos taurus opioid binding protein/cell adhesion molecule (OBCAM) mRNA (gb:GENBANK-ID:BTOBCAM|acc:X12672.1) (E = 1.3e^"56). A disclosed NOV5c polypeptide (SEQ ID NO:26) encoded by SEQ ID NO:25 is 307 amino acid residues and is presented using the one-letter code in Table 5F. Signal P, Psort and/or Hydropathy results predict that NOV5c does not contain a signal peptide and is likely to be localized in the mitochondrial matrix space with a certainty of 0.4686 and the cytoplasm with a certainty of 0.4500. Although SignalP, Psort and/or hydropathy suggest that NOV5c protein may be localized in the mitochondrial matrix space and cytoplasm, the protein predicted here is similar to the Opioid Binding Cell Adhesion Molecule family, some members of which are released extracellularly. The closest homolog SWISSPROT-ACC:Q14982 opioid binding protein/cell adhesion molecule is a type la membrane protein that is localized to the plasma membrane extracellularly. This indicates that the signal peptide ofthe mature protein is cleaved. Therefore it is likely that NOV5c protein is available at the same sub-cellular localization and hence accessible to a diagnostic probe and for various therapeutic applications. It also indicates that the use of a heterologous signal peptide to target the novel protein to the appropriate location, i.e. extracellularly, is appropriate.

Table 5F. Encoded NOV5c protein sequence (SEQ ID NO:26)

SQSLEFNSPADNYTVCEGDNATLSCFIDEHVTRVAWLNRSNILYAGNDRWTSDPRVWLLINTPEEFSILITEVGLGDEGLY TCSFQTRHQPYTTQVYLIVHVPARIVNISSPVMVNEGGNVNLLCLAVGRPEPTVTWRQLRDGFTSEGEILEISDIQRGQAG EYECVTHNGVNSAPDSRRVLVTVNYPPTITDVTSARTALGRAALLRCEAMAVPPADFQWYKDDRLLSSGTAEGLKVQMERT RSMLLFANVSARHYGNYTCRAANRLGASSASMRLLRPGSLENSAPRPPGLLALLSALGWLWWRM

The NOV5c amino acid sequence has 163 of 300 amino acid residues (54%) identical to, and 212 of 300 amino acid residues (70%) similar to, a Homo Sapiens 345 amino acid residue opioid binding protein/cell adhesion molecule precursor (OBCAM) / opioid-binding cell adhesion molecule (OPCML) protein (ptnr:SWISSPROT-ACC:Q14982) (E = 1.2e^"79). NOV5c is expressed in at least the following tissues: Brain and Fetal brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, genomic clone sources, literature sources, and/or RACE sources.

NOV5d

A disclosed NOV5d nucleic acid of 1169 nucleotides (also referred to as CG51027-05) encoding a novel Opioid Binding Cell Adhesion Molecule- like protein is shown in Table 5G. An open reading frame was identified beginning with an ATG codon at nucleotides 71-73 and ending with a TAG codon at nucleotides 1079-1081. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 5G, and the start and stop codons are in bold letters.

Table 5G. NOV5d Nucleotide Sequence (SEQ ID NO:27)

CTCC7ΛACAAGCATΛGCGCGCCCCGGΛCCGGCCCCCCCTTTCCCCTCCCCCTCCGTGCCGCCTCTGCCGCGATG CCCCCCGCTGCGCCCGGGGCCCGGCTCCGGCTTCTCGCCGCCGCCGCCCTGGCCGGCTTGGCCGTCATCAGCC GGGGGCTGCTCTCCCAGAGGCTGGAGTTCAACTCTCCTGCCGACAACTACACAGTGTGTGAAGGTGACAACGC CACCCTCAGCTGCTTCATGGACGAGCATGTGACCCGCGTGGCCTGGCTGAACCGCTCCAACATCCTGTACGCC GGCAACGACCGCAGGACCAGGGACCCGCGGGTGCGGCTGCTCATCAACACCCCCGAGGAGTTCTCCATCCTCG TCACCGAGGTGGGGCTCGGCGACGAGGGCCTCTACACCTGCTCCTTCCAGACCCGCCACCAGCCGTACACCAC TCAGGTCTACCTCATTGTCCACGTCCCTGCCCGCGTTGTGAACATCTCGTCGCCTGTGATGGTGAATGAGGGA GGTAATGTGAACCTGCTTTGCCTGGCCGTGGGGCGGCCAGAGCCCACGGTCACCTGGAGACAGCTCCGAGACG GCTTCACCTCGGAGGGAGAGATCCTGGAGATCTCTGACATCCTGCGGGGCCAGGCCGGGGAGTATGAGTGCGT GACTCACAACGGGGTTAACTCGGCGCCCGACAGCCGCCGCGTGCTGGTCACAGTCAACTATCCTCCGACCATC ACGGACGTGACCAGCGCCCGCACCGCGCTGGGGCCGGGCCGCCTACTGCGCTGCGAAGCCATGGCGGTTTCCC CCGCGGATTTCCAGTGGTATAAGGATGACAGACTACTGAGCAGCGGCACGGCCGAGGGCCTGAAGGTGCAGAT GGAGCGCACTCGCTCGATGCTTCTCTTTGCCAACATGAGCGCCCGGCATTACGGCAACTATACGTGTTGCGCC GCCAACCGGCTGGGAGCGTCCAGCGCCTCCATGCGGCTCCTGTGCCCAGGATCCCTGGAGAACTCAGCCCCGA GGCCCCCAGGGCCCCTGGCCCTCCTCTCCGCCCTGGGCTGGCTGTGGTGGAGAATGTAGGCGCAACCCAGTGG AGCTCGCCTCCCCCTGCAGGGGGCCTCAGGCCAAGAGTGAGAGAAAAGGGGGAGCAAGAGCCCTGGGTCTCGT

The NOV5d nucleic acid was identified on chromosome 7 and has 257 of 389 bases (66%) identical to a Homo sapiens (clone pHOM) opioid-binding cell adhesion molecule mRNA (gb:GENBANK-ID:HUMOBCAM|acc:L34774.1) (E = 2.0e^"46).

A disclosed NOV5d polypeptide (SEQ ID NO:28) encoded by SEQ ID NO:27 is 336 amino acid residues and is presented using the one-letter code in Table 5H. Signal P, Psort and/or Hydropathy results predict that NOV5d contains a signal peptide and is likely to be localized extracellularly with a certainty of 0.6329. The most likely cleavage site for a NOV5d peptide is between amino acids 30 and 31, at: LLS-QR.

Table 5H. Encoded NOV5d protein sequence (SEQ ID NO:28)

MPPAAPGARLRLLAAAALAGLAVISRGLLSQRLEFNSPADNYTVCEGDNATLSCFMDEHVTRVAWLNRSNIL YAGNDRRTRDPRVRLLINTPEEFSILVTEVGLGDEGLYTCSFQTRHQPYTTQVYLIVHVPARVVNISSPVMV NEGGNVNLLCLAVGRPEPTVTWRQLRDGFTSEGEILEISDILRGQAGEYECVTHNGVNSAPDSRRVLVTVNY PPTITDVTSARTALGPGRLLRCEAMAVSPADFQWYKDDRLLSSGTAEGLKVQMERTRSMLLFANMSARHYGN YTCCAANRLGASSASMRLLCPGSLENSAPRPPGPLALLSALGWLWWRM The NOV5d amino acid sequence has 160 of 327 amino acid residues (48%) identical to, and 218 of 327 amino acid residues (66%) similar to, a Bos taurus 345 amino acid residue opioid binding protein/cell adhesion molecule precursor (OBCAM) / opioid-binding cell adhesion molecule (OPCML) protein (ptnr:SWISSNEW-ACC:P11834) (E = 1.5e^"75).

NOV5d is expressed in at least the following tissues: Brain and Fetal brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, genomic clone sources, literature sources, and/or RACE sources. In addition, NOV5d is predicted to be expressed in occipital cortex tissues because ofthe expression pattern of a closely related Homo sapiens (clone pHOM) opioid-binding cell adhesion molecule mRNA homolog (GENBANK-ID: gb:GENBANK-ID:HUMOBCAM|acc:L34774.1).

Possible small nucleotide polymorphisms (SNPs) found for NOV5c are listed in Table 51.

NOV5a - NOV5d are very closely homologous as is shown in the amino acid alignment in Table 5J.

Table 5J Amino Acid Alignment of NOV5a and NOV5b

10 20 30 40 50 • I .. • I ..

NOV5a LAGLAVISRGLLSQSLEFNSPADNYTVCEGDNA NOV5b LAGLAVISRGLLSOSLEFNSPADNYTVCEGDNA NOV5c NOV5d MPPAAPGART.RT.T.AAAAIMWIKWMMaMIIII

110 120 130 140 150 . . I . . . . I . . . . | . . . . I

NOV5a ΪVGLGDEGLYTCSFQTRHQPYTTQVYLIVHVPARIVNISSPVJSVNEGGN\ NOV5b 2VGLGDEGLYTCSFOTRHQPYTTQVYLIVHVPARIVNISSPVHVNEGGN\ NOV5c ΞVGLGDEGLYTCSFQTRHQPYTTQVYLIVHVPARIVNISSPVIFFLVNEGGNVL NOV5d SVGLGDEGLYTCSFQTRHOPYTTQVYLIVHVPARSVNISSPVBVNEGGNV

210 220 230 240 250

Homologies to any ofthe above NOV5 proteins will be shared by the other NOV5 proteins insofar as they are homologous to each other as shown above. Any reference to NOV5 is assumed to refer to both ofthe NOV5 proteins in general, unless otherwise noted.

NOV5a also has homology to the amino acid sequences shown in the BLASTP data listed in Table 5K.

Table 5K. BLAST results for NOV5a

Gene Index/ Protein/ Organism Length Identity Positives Expect Identifier (aa) (%) (%)

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 5L.

Table 5L Clustal W Sequence Alignment

1) NOV5a (SEQ ID NO:22)

2) gi|129173|sp|Pl 1834IOPCM BOVIN opioid binding protein cell adhesion molecule precursor (OBCAM) (opioid-binding cell adhesion molecule) (OPCML) [Bos taurus]

(SEQ ID NO:83)

3) gi|4505505|reflNP 002536.1| (NM_002545) opioid-binding cell adhesion molecule precursor; opiate binding-cell adhesion molecule [Homo sapiens] (SEQ ID NO: 84)

4) gill352640lsplP32736IOPCM_RAT opioid binding protein/cell adhesion molecule precursor (OBCAM) (opioid-binding cell adhesion molecule) (OPCML) [Rattus norvegicus] (SEQ ID NO:85)

5) gi|112102|pir||JC1238 opioid-binding protein (clone DUZ1) [Rattus norvegicus] (SEQ ID NO:86)

6) gil3298456ldbilBAA31514.11 (AB011810) CEPU-1 [Gallus gallus] (SEQ ID NO:87)

260 270 280 290 300

310 320 330 340

Tables 5M - 5R list the domain description from DOMAIN analysis results against NOV5a. This indicates that the NOV5a sequence has properties similar to those of other proteins known to contain these domains.

Table 5M Domain Analysis of NOV5a gnl I Smart I smart00409, IG, Immunoglobulin (SEQ ID NO: 88)

Length = 86 residues, 100.0% aligned

Score = 53.1 bits (126), Expect = 2e-08

NOV5a 122 SSPVTVNEGGNVNLLCLAVGRPEPTVT RQ LRDGFTSEGE ILEISDIQR 170

I I I I I +1 I I I I I I I I I I + 1+ I I I++

00409 1 PPSVTVKEGESVTLSCEASGNPPPTVT YKQGGKLLAESGRFSVSRSGGNSTLTISNVTP 60

NOV5a 171 GQAGEYECVTHNGVNSAPDSRRVLVT 196

+1 1 1 I II + 1 00409 61 EDSGTYTCAATNSSGSASSGTTLTVL 86

Table 5N Domain Analysis of NOV5a gnl I Smart | smart00409, IG, Immunoglobulin (SEQ ID NO: 89)

Length = 86 residues, 87.2% aligned

Score = 49.3 bits (116), Expect = 3e-07

NOV5a 214 GRTALLRCEAMAVPPADFQ YKDDRLLSSGTAEGLKVQTERTRSMLLFANVSARHYGNYT 273 I + I I I I II III I + + I 1 1 +11+ I II 00409 9 GESVTLSCEASGNPPPTVT YKQGGKLLAES-GRFSVSRSGGNSTLTISNVTPEDSGTYT 67

NOV5a 274 CRAANRLGASSASMRL 289

I I I I++I+ I 00409 68 CAATNSSGSASSGTTL 8

Table 5O Domain Analysis of NOV5a gnl I Smart I smart00409, IG, Immunoglobulin (SEQ ID NO: 90)

Length = 86 residues, 97.7% aligned

Score = 43.9 bits (102) , Expect = le-05

NOV5a 24 NYTVCEGDNATLSCFIDEHVT-RVA LNRSNILYAGNDRWTSDPRVRLLINTPEEFSILI 82

+ I I I I++ I I I I + 1 1 + I I + I++ + ++ I

00409 3 SVTVKEGESVTLSCEASGNPPPTVTWYKQGGKLLAESGRFSVS RSGGNSTLTI 55

NOV5a 83 TEVGLGDEGLYTCSFQTRHQPYTTQVYLIVH 113

+ 1 I I II I + ++ I I 00409 56 SNVTPEDSGTYTCAATNSSGSASSGTTLTVL 86

Table 5P Domain Analysis of NOV5a gnl I Smart | smart00408, IGc2, Immunoglobulin C-2 Type (SEQ ID NO: 91)

Length = 63 residues, 100.0% aligned

Score = 51.6 bits (122) , Expect = 7e-08

NOV5a 128 NEGGNVNLLCLAVGRPEPTVT R QLRDG-FTSEGEILEISDIQRGQAGEYECVTHN 182

I I +1 I I I I I I +1 I 1 + + 1 I I ++ +1 I I I I 00408 1 LEGESVTLTCPASGDPVPNIT LKDG PLPESRVVASGSTLTIKNVSLEDSGLYTCVARN 60

NOV5a 183 GVN 185

I 00408 61 SVG 63

Table 5Q Domain Analysis of NOV5a gnl I Smart | smart00408, IGc2, Immunoglobulin C-2 Type (SEQ ID NO: 92)

Length = 63 residues, 96.8% aligned

Score = 50.8 bits (120) , Expect = le-07

NOV5a 214 GRTALLRCEAMAVPPADFQWYKDDRLLSSGTAEGLKVQTERTRSMLLFANVSARHYGNYT 273 l + l l l l + l l l + l + + I I I I I I I I

00408 3 GESVTLTCPASGDPVPNIT LKDGKPLPES RWASGSTLTIKNVSLEDSGLYT 55

NOV5a 274 CRAANRLG 281

I I I +1 00408 56 CVARNSVG 63

Table 5R Domain Analysis of NOV5a gnl I Pfam|pfam00047, ig, Immunoglobulin domain. Members of the immunoglobulin superfamily are found in hundreds of proteins of different functions. Examples include antibodies, the giant muscle kinase titin and receptor tyrosine kinases. Immunoglobulin-like domains may be involved in protein-protein and protein-ligand interactions. The Pfam alignments do not include the first and last strand of the immunoglobulin-like domain. (SEQ ID NO: 93) Length = 68 residues, 100.0% aligned Score = 35.0 bits (79), Expect = 0.007 N0V5a 214 GRTALLRCEAMAVPP-ADFQ YKDDRLLSSGTAEGLKVQTERTRSM LLFANVSARH 268

I + I I II I +I + + + +1 + 1+ I ++I+ 00407 1 GESVTLTCSVSGYPPDPTVTWLRDGKEIELLGSSESRVSSGGRFSISSLSLTISSVTPED 60

NOV5a 269 YGNYTCRA 276 l lll 00407 61 SGTYTCVV 68

Opioid-binding cell adhesion molecule (OBCAM), a neuron-specific protein, consists of three immunoglobulin (Ig)-like domains anchored to the membrane through a glycosylphosphatidylinositol (GPI)-tail. OBCAM has been presumed to play a role as a cell adhesion/recognition molecule, but its function has not been fully elucidated and may also play a role in early neuronal development (Hachisuka et al., Developmental expression of opioid-binding cell adhesion molecule (OBCAM) in rat brain. J Brain Res Dev Brain Res 122(2):183-91, 2000).

It has been previously reported that transfection of antisense OBCAM cDNA into NG108-15 neuroblastoma x glioma hybrid cells, which contain delta-opioid receptors, results in greatly reduced opioid binding (Ann et al., J. Biol. Chem. 267, 7921-7926, 1992) and more recently it has been reported that these cells show altered coupling between opioid receptors and G-proteins (Govitrapong et al., Transfection of NG108-15 cells with antisense opioid- binding cell adhesion molecule cDNA alters opioid receptor-G-protein interaction. J Biol Chem 268(24): 18280-5, 1993).

Despite the recent cloning of mu, delta and kappa opioid receptors, a role in opioid receptor function for OBCAM is supported by several lines of evidence, including inhibition of opioid binding by opioid binding cell adhesion molecule antibodies, down-regulation of opioid binding cell adhesion molecule by chronic opioid agonist treatment of cultured NG108- 15 cells, and reduction of opioid binding in NG108-15 cells by transfection of opioid binding cell adhesion molecule antisense cDNA (Kalyuzhny et al., An opioid binding protein is specifically down-regulated by chronic morphine treatment in dorsal root and trigeminal ganglia. Neuroscience 66(4):943-9, 1995).

The tissue distribution and brain localization of OBCAM was investigated in the adult rats. OBCAM was preferentially expressed in the central nervous system (CNS) and at a very low level in the spleen. Within the brain, OBCAM was distributed in almost all the gray matter, but little or no immunoreactive OBCAM was found in the white matter. Morphologically, the distribution pattern of OBCAM immunoreactivity was very similar to that of synaptophysin, suggesting a role in the synaptic machinery (Hachisuka et al., Localization of opioid-binding cell adhesion molecule (OBCAM) in adult rat brain.

842(2):482-6, 1999). Another study investigated the expression of two immunoglobulin superfamily (IgSF) proteins, Kilon and OBCAM. This study demonstrated the specific expression of Kilon and OBCAM in the hypothalamic magnocellular neurons, particularly in dendrites, suggesting that they confer on magnocellular neurons the ability to rearrange dendritic connectivity (Miyata et al., Expression ofthe IgLON cell adhesion molecules Kilon and OBCAM in hypothalamic magnocellular neurons. 424(l):74-85, 2000).

The above defined information for NOV5 suggests that this NON5 protein may function as a member of a Opioid Binding Cell Adhesion Molecule-like protein family. Therefore, the ΝOV5 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV5 compositions of the present invention will have efficacy for treatment of patients suffering from Von Hippel-Lindau (VHL) syndrome, Alzheimer's disease, stroke, tuberous sclerosis, hypercalceimia, Parkinson's disease, Huntington's disease, cerebral palsy, epilepsy, Lesch-Nyhan syndrome, multiple sclerosis, ataxia-telangiectasia, leukodystrophies, behavioral disorders, addiction, anxiety, pain, neurodegeneration. The NOV5 nucleic acid encoding Opioid Binding Cell Adhesion

Molecule-like protein, and the Opioid Binding Cell Adhesion Molecule- like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV6 NOV6 includes two novel triacylglycerol lipase-like proteins disclosed below. The disclosed proteins have been named NOV6a and NOV6b.

NOV6a

A disclosed NOV6a nucleic acid of 1377 nucleotides (also referred to as SCI 22982104_A) encoding a novel triacylglycerol lipase-like protein is shown in Table 6A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 91-93 and ending with a TAA codon at nucleotides 1243-1245. Putative untranslated regions upstream from the start codon and downstream from the termination codon are underlined in Table 6A, and the start and stop codons are in bold letters.

Table 6A. NOV6a Nucleotide Sequence (SEQ ID NO:29)

CAGTTTTATAAATGGTATTATTTGCTGCAATTTTAACTGTATΆAACATCTCCCTTTTATΆTATAGGTCCCAΆATGTGGTATC TTTTCACAATGATGTATTTTATCCGAATTCTTGGAATTACTCATGGTGTCTTTCAAAATTATAGATCTGTGAAACCTGAAGC AGATATGAATATTAGCCAGATTATTTCCTACTGGGGCTACCCTGATGAAGAATATGATATTGTAACCGAAGATGGTTATATC CTTGGCCTTTATAGAATTCCTTATTGGAGGACAGACAATAATAAAAATCTAGGTAATTCAGCTCAGAGGGTTGTTGTATACT TGCAACATGGTTTGCTTACATCTGCCAGCAGCTGGATTTCCAATCTTCCCAACAATAGTCTGGGCTTCATTCTGGCAGATGC TGGTTATGATGTGTGGATGGGAAATAGCAGAGGAAATACCTGGTCCAGGAAACACTTGTACCTAGAAACGAGTTCCAAAGAA TTCTGGGCTTTCAGGTACGCTCAAGGAGGCCTGCCTGCCTCTGTAGACTGCATCTTGGTGAAGAAGAGAGGAGAGAAAAATA TATATCATTATATCTTTCATTCCCAAGTACATAGCCAGGGAACCTTAGGTTTCATAACATTTTCTACTATATCAAAGATAGC

TGAAAGAATCAAAATATTTTTTGCTTTAGCACCCTCATCCTCAGTCAAATATACCAAGTCAATTATTCTTAAACTTACATAC AAATGGAAGTCAATAGGCAACAAAGACTTCTTGCCTAAAACCTCATTTAAAAAATTCATTGGTTCAAAGCTGTGTCCACTAC AGATTTTTGATAAGATTTGCCTCAATATCTTGTTTATGATGTTTGGATATGACCCAAAAAACTTAAATATGAGTCGTTTGGA TGTGTATTTTTCACACAACCCAGCAGGAACATCTGTTCAAAATATGCTTCATTGGAGTCAGGCTTATGACTGGGGCAGTCCT GATCTGAACTTGGTTCATTATAATCAGACAACGTCTCCATTATACAACATGACAAACATGAATGTGGCAACTGCAATTTGGA ATGGTAAAAGTGACTTGTTGGCTGACCCTGAAGACGTTAACATTTTACATTCTGAAATCACAAACCACATTTATTATAAAAC TATTTCTTACTACAATCATATAGACTCTTTGTTTGGATTAGATGTCTATGATCAAGTTTACCATGAAATCATTGATATTATC CAAGACΆATCTΆTAAAGΆACCΆTGGCGCTGTGTGTTTAAAGATCTACATCATTCCTAATGAAATCCAATTCTTATTTTTTTT TACCTGTGTATGTTCTTTCATTTTTAAAACTAAATATGTAGTTTTTTCCTCTATATTCTCATTGA

The NOV6a nucleic acid was identified on chromosome 10 and has 367 of 543 bases (67%) identical to a rabbit gastric lipase precursor coding sequence mRNA (gb:GENBANK- ID:A26689|acc:A26689.1) (E = 9.1e^"81).

A disclosed NOV6a polypeptide (SEQ ID NO:30) encoded by SEQ ID NO:29 is 390 amino acid residues and is presented using the one-letter code in Table 6B. Signal P, Psort and/or Hydropathy results predict that NOV6a contains a signal peptide and is likely to be localized in the microbody (peroxisome) with a certainty of 0.7480. The most likely cleavage site for a NOV6a peptide is between amino acids 19 and 20, at: THG-VF.

Table 6B. Encoded NOV6a protein sequence (SEQ ID NO:30)

MWYLFTMMYFIRILGITHGVFQNYRSVKPEAD NIΞQIISYWGYPDEEYDIVTEDGYILGLYRIPYWRTDNNKNLGNSAQR VVVYLQHGLLTSASSWISNLPNNSLGFILADAGYDV MGNSRGNT SRKHLYLETSSKEFWAFRYAQGGLPAΞVDCILVKK RGEKNIYHYIFHSQVHSQGTLGFITFSTISKIAERIKIFFALAPSSSVKYTKSIILKLTYK KSIGNKDFLPKTSFKKFIG SKLCPLQIFDKICLNILFMMFGYDPKNLNMSRLDVYFSHNPAGTSVQNMLHWSQAYDWGSPDLNLVHYNQTTSPLYNMTNM NVATAIWNGKSDLLADPEDVNILHSEITNHIYYKTISYYNHIDSLFGLDVYDQVYHEIIDIIQDNL

The NOV6a amino acid sequence has 218 of 396 amino acid residues (55%) identical to, and 288 of 396 amino acid residues (72%) similar to, a Homo sapiens 398 amino acid residue (triacylglycerol lipase, gastric precursor (ec 3.1.1.3) (gastric lipase) protein

(ptnr:SWISSPROT-ACC:P07098) (E = 8.0^"114). NOV6a is expressed in at least the following tissues: Whole Organism , fetal lung.

This information was derived by determining the tissue sources ofthe sequences that were included in the invention. SeqCalling sources: Whole Organism, PublicEST sources: fetal lung NbHL19W, testis NHT, and B-cell, pooled germ cell. In addition, NOV6a is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related precursor of rabbit gastric lipase coding sequence homolog in species synthetic construct (gb:GENBANK-ID:A26689|acc:A26689.1): lung, testis, tumors .

NOV6b

A disclosed NOV6b nucleic acid of 1260 nucleotides (also referred to as CG58608-02) encoding a novel triacylglycerol lipase-like protein is shown in Table 6C. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 31-33 and ending with a TAA codon at nucleotides 1234-1236. Putative untranslated regions upstream from the start codon and downstream from the termination codon are underlined in Table 6C, and the start and stop codons are in bold letters.

Table 6C. NOV6b Nucleotide Sequence (SEQ ID NO:31)

ATTAAATTTCCTTTCCTAGGCAGATCCCAAATGTGGCAGCTTTTAGCAGCAGCATGCTGGATGCTTCTTCTTGGATCTA TGTATGGTTATGACAAGAAAGGAAACAATGCAAACCCTGAAGCTAATATGAATATTAGCCAGATTATTTCCTACTGGGG CTACCCTGATGA_AGAATATGATATTGTAACCGAAGATGGTTATATCCTTGGCCTTTATAGAATTCCTTATTGGAGGACA GACAATAATAAAAATCTAGGTAATTCAGCTCAGAGGGTTGTTGTATACTTGCAACATGGTTTGCTTACATCTGCCAGCA GCTGGATTTCCAATCTTCCCAACAATAGTCTGGGCTTCATTCTGGCAGATGCTGGTTATGATGTGTGGATGGGAAATAG CAGAGGAAATACCTGGTCCAGGAAACACTTGTACCTAGAAACGAGTTCCAAAGAATTCTGGGCTTTCAGTTTTGATGAG ATGGCAAAATATGACCTTCCAGCCTCTATTGATTTCACTGTGAAGCAAACCAGACAAGAGGAAATATTTTATGTAGGCC ATTCACAGGGTACTACTATTGGTTTCATAACATTTTCTACTATATCAAAGATAGCTGAAAGAATCAAAATATTTTTTGC TTTAGCACCAGTTTTTTCCACAAAGTACTTAAAAAGTCCTTTAATTAGAATGACATACAAATGGAAGTCAATAGTCATG GCTTTTTCAGGCAACAAAGACTTCTTGCCTAAAACCTCATTTAAAAAATTCATTGGTTCAAAGCTGTGTCCACTACAGA TTTTTGATAAGATTTGCCTCAATATCTTGTTTATGATGTTTGGATATGACCCAAAAAACTTAAATATGAGTCGTTTGGA TGTGTATTTTTCACACAACCCAGCAGGAACATCTGTTCAAAATATGCTTCATTGGAGTCAGCTTTTAAATTCTACTCAT TTGAAAGCTTATGACTGGGGCAGTCCTGATCTGAACTTGGTTCATTATAATCAGACAACGTCTCCATTATACAACATGA CAAACATGAATGTGGCAACTGCAATTTGGAATGGTAAAAGTGACTTGTTGGCTGACCCTGAAGACGTTAACATTTTACA TTCTGAAATCACAAACCACATTTATTATAAAACTATTTCTTACTACAATCATATAGACTCTTTGTTTGGATTAGATGTC TATGATCAAGTTTACCATGAAATCΆTTGATATTATCCAAGACAATCTATAAAGAACCATGGCGCTGTGTGTTTAΆ

The NOV6b nucleic acid was identified on chromosome 1 and has 711 of 997 bases (71%) identical to a Homo sapiens gastric lipase mRNA (gb:GENBANK- ID:A01046|acc:A01046.1) (E = 1.3e^"117)

A disclosed NOV6b polypeptide (SEQ ID NO:32) encoded by SEQ ID NO:31 is 401 amino acid residues and is presented using the one-letter code in Table 6D. Signal P, Psort and/or Hydropathy results predict that NOV6b contains a signal peptide and is likely to be localized in the lysosome (lumen) with a certainty of 0.5078. The most likely cleavage site for a NOV6b peptide is between amino acids 19 and 20, at: MYG-YD.

Table 6D. Encoded NOV6b protein sequence (SEQ ID NO:32).

MWQLLAAACWMLLLGSMYGYDKKGNNANPEANMNISQIIS WGYPDEEYDIVTEDGYILGLYRIPYWRTDNNKNLGNSAQRVVVY LQHGLLTSASS ISNLPNNSLGFILADAGYDVWMGNSRGNTWSRKHLYLETSSKEFWAFSFDEMAKYDLPASIDFTVKQTRQEEI FYVGHSQGTTIGFITFSTISKIAERIKIFFALAPVFSTKYLKSPLIRMTYKWKSIVMAFSGNKDFLPKTSFKKFIGSKLCPLQIF DKICLNILFMMFGYDPKNLNMSRLDVYFSHNPAGTSVQNMLHWSQLLNSTHLKAYDWGSPDLNLVHYNQTTSPLYNMTNMNVATA IWNGKSDLLADPEDVNILHSEITNHIYYKTISYYNHIDSLFGLDVYDQVYHEIIDIIQDNL

The NOV6b amino acid sequence has 224 of 400 amino acid residues (56%) identical to, and 298 of 400 amino acid residues (74%) similar to, a Homo sapiens 398 amino acid residue (triacylglycerol lipase, gastric precursor (EC 3.1.1.3) (gastric lipase) protein (ptnr:SWISSPROT-ACC:P07098) (E = 3.5e^-124).

NOV6b is expressed in at least the following tissues: Mammalian Tissue. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Literature sources, and/or RACE sources.

Possible small nucleotide polymorphisms (SNPs) found for NOV6a are listed in Table 6E.

Table 6E: SNPs

NOV6a and NOV6b are very closely homologous as is shown in the amino acid alignment in Table 6F.

Table 6F Amino Acid Alignment of NOV6a and NOV6b

10 20 30 40 50 |...._ ...|....|....|....|.

NOV6a JJYJJFTMMYBΦRΪØBITHHVFINYRIVKI NOV6b |LAAACaML|JEsMg@YDKKGNNA |3SsH

60 70 80 90 100

. I.. • I.. .. I

NOV6a IVTEDGYILGLYRIPYWRTDNNKNLGNSAQRVVVYLQHGLLTSASSWISN NOV6b EVTEDGYILGLYRIPY RTDNNKNLGNSAORVVVYLQHGLLTSASSWISN

110 120 130 140 150 .1 • I.... I

NOV6a ■■■■■ϋl m ■ NOV6b aSrasraasFDEMS

210 220 230 240 250

260 270 280 290 300

..I .. ..|.. .. I

NOV6a IGSKLCPLQIFDKICLNILFM FGYDPKNLNMSRLDVYFSHNPAGTSVQN NOV6b IGSKLCPLQIFDKICLNILFMMFGYDPKNLNMSRLDVYFSHNPAGTSVON

310 320 330 340 350 ..I .. .. |.. .. I

NOV6a <.YDWGSPDLNLVHYNQ^r ΪPLYNMTNMNVATAIWNG NOV6b SLLNSTHL] AYDWGSPDLNLVHYNQ' 5PLYNMTNMNVATA1WNG

Homologies to any ofthe above NOV6 proteins will be shared by the other NOV6 proteins insofar as they are homologous to each other as shown above. Any reference to NOV6 is assumed to refer to both ofthe NOV6 proteins in general, unless otherwise noted.

NOV6a also has homology to the amino acid sequences shown in the BLASTP data listed in Table 6G.

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 6H.

Table 6H Information for the ClustalW proteins l) NOV6a (SEQ ID NO:30)

2) gil4758676lref1NP 004181.1| (NM_004190) lipase, gastric [Homo sapiens] (SEQ ID NO:94)

3) gi|758064|emb|CAA29414.11 (X05997) gastric lipase precursor [Homo sapiens] (SEQ ID NO:95)

4) gi|3041702|sp|P80035|LIPG_CANFA triacylglycerol lipase, gastric precursor (gastric lipase) (GL) [Canis familiaris] (SEQ ID NO:96)

5) gi|7546565|pdb|lHLG|A Chain A, Crystal Structure Of Human Gastric Lipase (SEQ ID NO:97)

6) gi|8394193|ref|NP 059037.1| (NM_017341) lingual lipase [Rattus norvegicus] (SEQ ID NO:98)

10 20 30 40 50

110 120 130 140 150

NOV6a DII HQΘ L- gi I 47586761 ISEBKK gi I 758064 I s|fflκκ gi j 3041702 I IGTHNK gi I 7546565 | ISEBKK gi I 8394193 I

The hydrolysis of triglycerides under influence of lipoprotein lipase is among the first recognised and well defined processes in postprandial lipid metabolism. More recently, also hepatic lipase has been implicated in the clearing of postprandial lipoproteins. Lipoprotein lipase as well as hepatic lipase are also involved in the metabolism of several other lipoproteins. However, their capacity is limited. This may lead to interaction of different metabolic processes and competition for the available lipase by different lipoproteins. Indeed, it is generally accepted that the exaggerated postprandial response in subjects with hypertriglyceridemia is at least partially due to competition between endogenous (VLDL) and exogenous (chylomicrons) lipoproteins. Similar mechanisms may also take place in the liver where hepatic lipase plays a role in the metabolism of several lipoproteins (Jansen et al., Role of lipoprotein lipases in postprandial lipid metabolism. Atherosclerosis 141 Suppl l :S31-4, 1998). Hepatic lipase (HL) is an enzyme that is made primarily by hepatocytes (and also found in adrenal gland and ovary) and hydrolyzes phospholipids and triglycerides of plasma lipoproteins. It is secreted and bound to the hepatocyte surface and readily released by heparin. It is a member ofthe lipase superfamily and is homologous to lipoprotein lipase and pancreatic lipase. The enzyme can be divided into an NH2 -terminal domain containing the catalytic site joined by a short spanning region to a smaller COOH-terminal domain. The NH2-terminal portion contains an active site serine in a pentapeptide consensus sequence, Gly-Xaa-Ser-Xaa- Gly, as part of a classic Ser-Asp-His catalytic triad, and a putative hinged loop structure covering the active site. The COOH-terminal domain contains a putative lipoprotein-binding site. The heparin-binding sites may be distributed throughout the molecule, with the characteristic elution pattern from heparin-sepharose determined by the COOH-terminal domain. Ofthe three N-linked glycosylation sites, Asn-56 is required for efficient secretion and enzymatic activity. HL is hypothesized to directly couple HDL lipid metabolism to tissue/cellular lipid metabolism. The potential significance ofthe HL pathway is that it provides the hepatocyte with a mechanism for the uptake of a subset of phospholipids enriched in unsaturated fatty acids and may allow the uptake of cholesteryl ester, free cholesterol, and phospho lipid without catabolism of HDL apolipoproteins. HL can hydro lyze triglyceride and phospholipid in all lipoproteins, but is predominant in the conversion of intermediate density lipoproteins to LDL and the conversion of post-prandial triglyceride-rich HDL into the postabsorptive triglyceride-poor HDL. HL plays a secondary role in the clearance of chylomicron remnants by the liver. A rare family with HL deficiency has been described. Affected patients are compound heterozygotes for a mutation of Ser267 to Phe that results in an inactive enzyme and a mutation of Thr383 to Met that results in impaired secretion and reduced specific activity. Human HL deficiency in the context of a second factor causing hyperlipidemia is strongly associated with premature coronary artery disease (Connelly and Hegele, Hepatic lipase deficiency. Crit Rev Clin Lab Sci 35(6):547-72, 1998).

The above defined information for NOV6 suggests that NOV6 may function as a member of a triacylglycerol lipase family. Therefore, the NOV6 nucleic acids and proteins of the invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV6 compositions of the present invention will have efficacy for treatment of patients suffering from Adrenoleukodystrophy , Congenital Adrenal Hyperplasia, Diabetes, Von Hippel-Lindau (VHL) syndrome , Pancreatitis, Obesity ,Endometriosis, Xerostomia, Scleroderma, Hypercalceimia, Ulcers Von Hippel-Lindau (VHL) syndrome, Cirrhosis, Transplantation, Inflammatory bowel disease, Diverticular disease, Hirschsprung's disease , Crohn's Disease, Appendicitis, Hypercalceimia, Arthritis, Ankylosing spondylitis, Arthritis, Tendinitis on Hippel-Lindau (VHL) syndrome , Alzheimer's disease, Stroke, Tuberous sclerosis, hypercalceimia, Parkinson's disease, Huntington's disease, Endocrine dysfunctions, Diabetes, Growth and reproductive disorders, Multiple sclerosis, Leukodystrophies, Pain, Myasthenia gravis,

Autoimmune disease, Asthma, Emphysema, Scleroderma, allergy, ARDS, Psoriasis, Actinic keratosis .Tuberous sclerosis, Acne, Hair growth, allopecia, pigmentation disorders, endocrine disorders Diabetes, Renal artery stenosis, Interstitial nephritis, Glomerulonephritis, Polycystic kidney disease, erythematosus, Renal tubular acidosis, IgA nephropathy, Hypercalceimia, Lesch-Nyhan syndrome. The NOV6 nucleic acid encoding triacylglycerol lipase-like protein, and the triacylglycerol lipase-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV7 NOV7 includes two novel IGE receptor beta subunit-like proteins disclosed below.

The disclosed proteins have been named NOV7a and NOV7b.

NOV7a

A disclosed NOV7a nucleic acid of 691 nucleotides (also referred to SC126624027_A) encoding a novel IGE receptor beta subunit-like protein is shown in Table 7A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 52-54 and ending with a TGA codon at nucleotides 652-654. Putative untranslated regions are found upstream from the initiation codon and downstream from the termination codon in Table 7A, and the start and stop codons are in bold letters.

Table 7A. NOV7a Nucleotide Sequence (SEQ ID NO:33)

GACTGAAGTACCAACTAAGTCATCTCCTTTCAAATTATCACCGACACCATCATGGATTCAAGCACCGCΆCACAGTCCGGT GTTTCTGGTATTTCCTCCAGAAATCACTGCTTCAGAATATGAGTCCACAGAACTTTCAGCCACGACCTTTTCAACTCAAA GCCCCTTGCAAAAATTATTTGCTAGAAAAATGAAAATCTTAGGGACTATCCAGATCCTGTTTGGAATTATGACCTTTTCT TTTGGAGTTATCTTCCTTTTCACTTTGTTAAAACCATATCCAAGGTTTCCCTTTATATTTCTTTCAGGATATCCATTCTG GGGCTCTGTTTTGTTCATTAATTCTGGAGCCTTCCTAATTGCAGTGAAAAGAAAAACCACAGAAACTCTGATAATATTGA GCCGAATAATGAATTTTCTTAGTGCCCTGGGAGCAATAGCTGGAATCATTCTCCTCACATTTGGTTTCATCCTAGATCAA AACTACATTTGTGGTTATTCTCACCAAAATAGTCAGTGTAAGGCTGTTACTGTCCTGTTCTTGGGAATTTTGATTACATT GATGACTTTCAGCATTATTGAATTATTCATTTCTCTGCCTTTCTCAATTTTGGGGTGCCACTCAGAGGATTGTGATTGTG AACAATGTTGTTGACTAGCACTGTGAGAATAAAGATGTGTTAAAATAAAAA

The disclosed NOV7a nucleic acid sequence, localized to chromosome 15, has 325 of 560 bases (58%) identical to a Mus musculus mast cell high affinity IgE receptor (Fc-epsilon- RI) beta subunit mRNA (gb:GENBANK-ID:MUSFCERB|acc:J05019.1) (E = 2.2e^"05). A disclosed NOV7a polypeptide (SEQ ID NO:34) encoded by SEQ ID NO:33 is 200 amino acid residues and is presented using the one-letter amino acid code in Table 7B. Signal P, Psort and/or Hydropathy results predict that NOV7a contains a signal peptide and is likely to be localized in the plasma membrane with a certainty of 0.6000. The most likely cleavage site for a NOV7a peptide is between amino acids 20 and 21, at: ITA-SE.

Table 7B. Encoded NOV7a protein sequence (SEQ ID NO:34).

MDSST_AHSP_VFL_VFPPEIT_ASEYESTELS_ATTFSTQSPLQKLFARKMKILGTIQILFGIMTFSFGVIFLFTLLKPYPRFP FIFLSGYPF_WGS_VLFI_NSG_AFLI_AVKRKTTETLIILSRIMNFLSALGAIAGIILLTFGFILDQNYICGYSHQNSQCKAVT VLFLGILITLMTFSIIELFISLPFSILGCHSEDCDCEQCC

The NOV7a amino acid sequence has 52 of 184 amino acid residues (28%) identical to, and 88 of 184 amino acid residues (47%) similar to the Homo Sapiens 214 amino acid residue IGE receptor beta subunit protein (ptnr:REMTREMBL-ACC:AAA62319) (E = 2.9e^"

¹³)-

NOV7a is expressed in at least the following tissues: Testis Whole Organism Male

Reproductive System; SeqCalling_celltypes: testis liver spleen parathyroid tumor. This information was derived by determining the tissue sources ofthe sequences that were included in the invention. SeqCalling sources: Testis Whole Organism Male Reproductive System,

PublicEST sources: testis liver spleen parathyroid_tumor. In addition, NOV7a is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus mast cell high affinity IgE receptor (Fc-epsilon-RI) beta subunit homolog mRNA

(GENBANK-ID: gb:GENBANK-ID:MUSFCERB |acc:J05019.1):Whole Organism Male

Reproductive System.

NOV7b

A disclosed NOV7b nucleic acid of 500 nucleotides (also referred to CG55760-02) encoding a novel IGE receptor beta subunit-like protein is shown in Table 7C. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 26-28 and ending with a TGA codon at nucleotides 473-475. Putative untranslated regions are found upstream from the initiation codon and downstream from the termination codon in Table 7C, and the start and stop codons are in bold letters.

Table 7C. NOV7b Nucleotide Sequence (SEQ ID NO:35)

CTTTCAAATTATCACCGACACCATCATGGATTCAAGCACCGCΆCACAGTCCGGTGTTTCTGGTATTTCCTCCAGAAATCA CTGCTTCAGAATATGAGTCCACAGAACTTTCAGCCACGACCTTTTCAACTCAAAGCCCCTTGCAAAAATTATTTGCTAGA AAAATGAAAATCTTAGGGACTATCCAGATCCTGTTTGGAATTATGACCTTTTCTTTTGGAGTTATCTTCCTTTTCACCTT GTTAAAACCATATCCAAGGTTTCCCTTTATATTTCTTTCAGGATATCCATTCTGGGGCTCTGTTTTGTTCATTAATTCTG GAGCCTTCCTAATTGCAGTGAAAAGAAAAACCACAGAAACTCTGGGAATTTTGATTACATTGATGACTTTCAGCATTATT GAATTATTCATTTCTCTGTCTTTCTCAΆTTTTGGGGTGCCACTCAGAGGATTGTGATTGTGAACΆATGTTGTTGACTAGC ACTGTGAGAATAAAGATGTG The disclosed NOV7b nucleic acid sequence, localized to chromosome 15, has 167 of 269 bases (62%) identical to a Mus musculus mast cell high affinity IgE receptor (Fc-epsilon- RI) beta subunit mRNA (gb:GENBANK-ID:MUSFCERB|acc:J05019.1) (E = 0.0023).

A disclosed NOV7b polypeptide (SEQ ID NO: 36) encoded by SEQ ID NO:35 is 149 amino acid residues and is presented using the one-letter amino acid code in Table 7D. Signal P, Psort and/or Hydropathy results predict that NOV7b contains a signal peptide and is likely to be localized in the plasma membrane with a certainty of 0.6000. The most likely cleavage site for a NOV7b peptide is between amino acids 20 and 21, at: ITA-SE.

Table 7D. Encoded NOV7b protein sequence (SEQ ID NO:36).

MDSΞTAHSPVFLVFPPEITASEYESTELSATTFSTQSPLQKLFARKMKILGTIQILFGIMTFSFGVI FLFTLLKPYPRFP FI FLSGYPFWGSVLFINSGAFLIAVKRKTTETLGILITLMTFSI IELFI SLSFSILGCHSEDCDCEQ.EE

The NOV7b amino acid sequence has 117 of 122 amino acid residues (95%) identical to, and 118 of 122 amino acid residues (96%) similar to the Homo Sapiens 200 amino acid residue MS4A5 protein (ptnr:TREMBLNEW-ACC:BAB18739) (E = 5.7e^"57).

NOV 7b is expressed in at least the following tissues: testis. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Literature sources, and/or RACE sources.

Possible small nucleotide polymorphisms (SNPs) found for NOV7a are listed in Table 7E.

NOV7a and NOV7b are very closely homologous as is shown in the amino acid alignment in Table 7F.

Table 7F Amino Acid Alignment of NOV7a and NOV7b

10 20 30 40 50 I I

NOV7a MDSSTAHSPVFLVFPPEITASEY NOV7b MDSSTAHSPVFLVFPPEITASEY

60 70 30 90 100

NOV7a GTIQILFGIMTFSFGVIFLFTLLKPYPRFPFIFLSGYPFWGSVLFINSGA NOV7b GTIQILFGIMTFSFGVIFLFTLLKPYPRFPFIFLSGYPFWGSVLFINSGA

110 120 130 140 150

. . I . . . . I . . . . I . . . . I . . . . | . . . . | . . . . | . . . . | . . . . | NOV7a FLIAVKRKTTET I I ILSRIMNFLSALGAIAGI ILLTFGFILDQNYICGYS NOV7b FLIAVKRKTTET

160 170 180 190 200

NOV7a HQNSQCKAVTVLEfi NOV7b mmπππ

Homologies to any ofthe above NOV7 proteins will be shared by the other NOV7 proteins insofar as they are homologous to each other as shown above. Any reference to NOV7 is assumed to refer to both ofthe NOV7 proteins in general, unless otherwise noted.

NOV7a also has homology to the amino acid sequence shown in the BLASTP data listed in Table 7G.

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 7H.

Table 7H. Information for the ClustalW proteins

1) NOV7a (SEQ ID NO:34)

2) gi|13647408|ref1XP 015541.1 | (XM_015541) hypothetical protein XP_015541 [Homo sapiens] (SEQ ID NO:99)

3) gi|12965205|ref1NP_076434.1| (NM_023945) testis-expressed transmembrane-4 protein [Homo sapiens] (SEQ ID NO: 100)

4) gi|16184634lref]XP 043510.21 (XM_043510) hypothetical protein XP_043510 [Homo sapiens] (SEQ ID NO:101)

5) gill4517598|dbj|B AB61018.il (AB022821) similar to Fc epsilon receptor beta subunit [Homo sapiens] (SEQ ID NO: 102)

6) gill6184624lref1XP 015539.31 (XM_015539) hypothetical protein XP_015539 [Homo sapiens] (SEQ ID NO: 103)

gi|16184624| - -MJ|TMQG|EQAMBGAGPGVPQLGNMAVIH|HL

60 70 80 90 100

110 120 130 140 150

The IgE receptor plays a central role in allergic disease, coupling allergen and mast cell to initiate the inflammatory and immediate hypersensitivity responses that are characteristic of disorders such as hay fever and asthma. The allergic response occurs when 2 or more high- affinity IgE receptors are crosslinked via IgE molecules that in turn are bound to an allergen (antigen) molecule. A perturbation occurs that brings about the release of histamine and proteases from the granules in the cytoplasm ofthe mast cell and leads to the synthesis of prostaglandins and leukotrienes-potent effectors ofthe hypersensitivity response. The IgE receptor consists of 3 subunits: alpha, beta (147138), and gamma (147139); only the alpha subunit is glycosylated. Shimizu et al. (1988) cloned and sequenced cDNAs for the rat and human alpha subunits of high-affinity IgE receptor. Both encode an NH2-terminal signal peptide, 2 immunoglobulin-like extracellular domains (encoded by discrete exons), a hydrophobic transmembrane region, and a positively charged cytoplasmic tail. The human and rat alpha subunits share similarities with each other and with the immunoglobulin gene family, suggesting origin from a common ancestral gene, and share structural homology with their ligands. Liu et al. (1988) used a synthetic oligonucleotide homologous to the amino-terminal sequence ofthe alpha subunit to screen a cDNA library from a rat basophilic leukemia cell line. Nucleotide sequencing demonstrated 4 distinct varieties of cloned cDNAs, differing at the 5 -prime ends and within the region encoding the second extracellular domain, suggesting the existence of at least 4 distinct protein products ofthe gene. By study of somatic cell hybrid DNA and in situ hybridization, Tepler et al. (1989) assigned the alpha chain ofthe Fc IgE receptor to human Iq21-q23. Seldin (1989) assigned the homologous mouse gene to chromosome 1. By in situ hybridization, Le Coniat et al. (1990) assigned genes for both the alpha and the gamma subunits to lq23. Garman et al. (1998) determined the x-ray crystal structure ofthe antibody-binding domains ofthe human IgE receptor alpha subunit at 2.4- angstrom resolution. The structure revealed a highly bent arrangement of immunoglobulin domains that form an extended convex surface of interaction with IgE. A prominent loop that confers specificity for IgE molecules extends from the receptor surface near an unusual arrangement of 4 exposed tryptophans. The crystal structure ofthe IgE receptor provides a foundation for the development of new therapeutic approaches to allergy treatment. Garman et al. (2000) solved the crystal structure ofthe human IgE-Fc-FCERl A complex to 3.5-angstrom resolution. The crystal structure revealed that 1 receptor binds 1 dimeric IgE-Fc molecule asymmetrically to interactions at 2 sites, each involving 1 C-epsilon-3 domain ofthe IgE-Fc. The interaction of 1 receptor with the IgE-Fc blocks the binding of a second receptor, and features of this interaction are conserved in other members ofthe Fc receptor family. (Garman et al., Crystal structure ofthe human high-affinity IgE receptor. Cell 95: 951-961, 1998; Garman et al., Structure ofthe Fc fragment of human IgE bound to its high-affinity receptor Fc-epsilon-RI-alpha. Nature 406: 259-266, 2000; Le Coniat et al., The human genes for the alpha and gamma subunits ofthe mast cell receptor for immunoglobulin E are located on human chromosome band lq23. Immunogenetics 32: 183-186, 1990; Liu et al., cDNA heterogeneity suggests structural variants related to the high-affinity IgE receptor. Proc. Nat. Acad. Sci. 85: 5639-5643, 1988; Shimizu et al., Human and rat mast cell high-affinity immunoglobulin E receptors: characterization of putative alpha-chain gene products. Proc. Nat. Acad. Sci. 85: 1907-1911, 1988; Tepler et al., The gene for the human mast cell high- affinity IgE receptor alpha chain: chromosomal localization to Iq21-q23 and RFLP analysis. Am. J. Hum. Genet. 45: 761-765, 1989).

The above defined information for NOV7 suggests that this NOV7 protein may function as a member of a IGE receptor beta subunit protein family. Therefore, the NOV7 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV7 compositions ofthe present invention will have efficacy for treatment of patients suffering from inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; blood disorders; asthma; psoriasis; inflammatory skin disordersvascular disorders, hypertension, skin disorders, renal disorders including Alport syndrome, immunological disorders, inflammation including irritable bowel disease, and tissue injury, cancers, fibrosis disorders, bone diseases, Ehlers-Danlos syndrome type VI, VII, type IV, S-linked cutis laxa and Ehlers- Danlos syndrome type V, osteogenesis imperfecta.Immuno therapy of inflammatory and infectious diseases such as AIDS, cancer therapy, treatment of Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), anorexia, bulimia, asthma, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, Crohn's disease; multiple sclerosis; and Treatment of Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, asthma, allergies, benign prostatic hypertrophy, and psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease or Gilles de la Tourette syndrome. The NOV7 nucleic acid encoding IGE receptor beta subunit-like protein, and the IGE receptor beta subunit-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV8

A disclosed NOV8 nucleic acid of 1386 nucleotides (also referred to SC138745558_A) encoding a novel Muncl8-like protein is shown in Table 8 A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 36-38 and ending with a TGA codon at nucleotides 1350-1352. Putitive untranslated regions upstream from the initiation codon and downstream from the termination codon are underlined in Table 8A, and the start and stop codons are in bold letters.

Table 8A. NOV8 Nucleotide Sequence (SEQ ID NO:37)

CACTCTGGCCGTGGCCGACGTTGCTCCTCCGAAGCATGGCGCCGGCGGGAGTTGTAACCCGGGCTGTCCGGAGCGGGGAG CTGCCCCTCACAAGCATGGCGTCAGCTGAAAATGAAGCCTGTGCTGTGCGGAGCGTCGCCTGCCCCTCACAAGCATGGCG TCTGCAGAAGGTGCTGTGCGGCCGCTGCGGGGCGGCCAGCTGCCCTTCACAAACATGGCGGCCGAGGGGTGCGGGGAGTG GCGGGGTAAGGATGGGAAGCCGAGCAGACGGCCCCAGAACAAGCGGTCATGTGACTGGGAAGATGGCCGTCTTTCCTTGG CACTCCAGGAATAGGAACTACAAAGCTGAATTTGCATCATGCCGACTGGAGGCTGTACCATTGGAGTTTGGGGACTATCA CCCTCTGAAACCCATAACTGTCACAGAGTCAAAGACAAAGAAAGTGAACCGGAAAGGAAGCACTTCTTCCACGTCCTCCT CCTCCTCCAGCTCCGTGGTGGACCCGCTGAGCAGCGTCCTCGATGGGACTGACCCCCTCTCCATGTTTGCAGCCACTGCT GACCCCGCAGCCTTGGCAGCTGCCATGGACAGCTCCAGAAGGAAACGTGATAGAGATGATAACTCCGTTGTAGGATCGGA TTTTGAGCCTTGGACCAACAAACGGGGAGAAATCCTTGCCCGGTACACCACTACCGAAAAGCTGTCTATTAATCTGTTTA TGGGATCTGAAAAAGGCAAAGCTGGGACTGCCACATTGGCAATGTCAGAGAAGGTGCGGACCCGGCTGGAGGAGCTGGAT GACTTTGAGGAGGGTTCCCAAAAGGAGCTGTTGAACTTGACTCAGCAGGATTACGTGAACCGCATAGAGGAGCTCAACCA ATCGCTGAAGGATGCCTGGGCCTCAGACCAGAAAGTGAAGGCTCTAAAAATAGTCATCCAGTGTTCAAAGCTTCTTTCAG ACACCAGTGTTATTCAGTTCTACCCAAGCAAATTTGTCCTTATCACCGACATACTTGATACATTTGGAAAGCTCGTGTAC GAGCGCATCTTTTCCATGTGTGTGGATAGCCGCAGCGTCTTACCAGATCACTTTTCTCCAGAGAATGCAAATGACACGGC CAAGGAAACATGCCTAAATTGGTTTTTCAAGATTGCCTCCATCAGGGAACTCATTCCAAGATTTTACGTGGAGGCATCCA TCCTGAAATGTAACAAATTCCTCTCCAAAACGGGAATTTCAGAGTGCCTGCCCCGGTTGACATGCATGATCAGAGGGATC GGAGACCCACTAGTGTCGGTGTATGCCCGTGCCTACCTGTGCCGGGTAGGCCATGCGAGTCACTGCCCTTGACGGCACGG CTGCCACCCGTCTCCTTGAATGTTCC

The disclosed NOV8 nucleic acid sequence is localized to chromosome 16.

A disclosed NOV8 polypeptide (SEQ ID NO:38) encoded by SEQ ID NO:37 is 438 amino acid residues and is presented using the one-letter amino acid code in Table 8B. Signal P, Psort and/or Hydropathy results predict that NOV8 does not contain a signal peptide and is likely to be localized to the mitochondrial matrix space with a certainty of 0.4363.

Table 8B. Encoded NOV8 protein sequence (SEQ ID NO:38).

MAPAGVVTRAVRSGELPLTSMASAENEACAVRΞVACPSQAWRLQKVLCGRCGAASCPSQTWRPRGAGSGGVRMGSRADGP RTSGHVTGKMAVFPWHSRNRNYKAEFASCRLEAVPLEFGDYHPLKPITVTESKTKKVNRKGΞTSSTSSSSSSSVVDPLSS VLDGTDPLSMFAATADPAALAAAMDSSRRKRDRDDNSVVGSDFEPWTNKRGEILARYTTTEKLSINLFMGSEKGKAGTAT LAMSEKVRTRLEELDDFEEGSQKELLNLTQQDYVNRIEELNQSLKDAWASDQKVKALKIVIQCSKLLSDTSVIQFYPSKF VLITDILDTFGKLVYERIFSMCVDSRSVLPDHFSPENANDTAKETCLNWFFKIASIRELIPRFYVEASILKCNKFLSKTG ISECLPRLTC IRGIGDPLVSVYARAYLCRVGHASHCP

The NOV8 amino acid sequence has 204 of 204 amino acid residues (100%) identical to, and 204 of 204 amino acid residues (100%) similar to, a Homo sapiens 824 amino acid residue FLJ21040 FIS, CLONE CAE 10642 protein (ptnr:TREMBLNEW-ACC:BAB14965)(E = 1.9e^"105).

NOV8a is expressed in at least the following tissues: Adrenal Gland/Suprarenal gland, Amygdala, Cervix, Pituitary Gland, Thymus, Tonsils, Whole Organism, SeqCalling_celltypes: liver, spleen, testis, tumor. This information was derived by determining the tissue sources of the sequences that were included in the invention. SeqCalling sources: Adrenal

Gland/Suprarenal gland, Amygdala, Cervix, Pituitary Gland, Thymus, Tonsils, Whole Organism; PublicEST sources: liver, spleen, testis, tumor.

The disclosed NOV8 polypeptide has homology to the amino acid sequences shown in the BLASTP data listed in Table 8C.

The homology between these and other sequences is shown graphically in the ClustalW analysis shown in Table 8D.

Table 8D. ClustalW Analysis of NOV8

1) Novel NOV8 (SEQ ID NO:38)

2) gill2858347ldbj|B AB31285.il (AK018573) putative [Mus musculus] (SEQ ID NO: 104)

3) gi|14777378|ref1XP 040052.11 (XM_040052) hypothetical protein XP_040052 [Homo sapiens] (SEQ ID NO: 105)

4) gi|12847855[dbj|BAB27735.1| (AK011615) putative [Mus musculus] (SEQ ID NO:106)

5) gi| 15292597]gb|AAK93567.11 (AY052143) SD10311p [Drosophila melanogaster] (SEQ ID NO: 107)

6) gi|7299081 |gblAAF54281.11 (AE003680) CG8202 gene product [Drosophila melanogaster] (SEQ ID NO: 108)

10 20 30 40 50 I I I I . . . . I I I . . . . I I I

NOV8 APAGVVTRAVRΞGELPLTSMASAENEACAVRSVACPSQAWRLQKVLCGR gi 112858347 | gi 114777378 | gi 112847855 I gil 15292597 I gi I 72990811

60 70 80 90 100

— I — i — i — i — i — i . . . . i — j_. ■ • I

110 120 130 140 150

1 m m mm mm H s* |QR0V|VVDB- gil 7299081 | DNNNHD0MAN 5cgPR--CJfEV KNcl0GQABL|HPL

160 170 180 190 200

310 320 330 340 350 ..| .. .. I ..

NOV8 JQKVKALKIVIQCSKLLSDTSVIQFYPSKFVLITDILDTFGKLVYI gi I 12858347 | JQKVKALKIVIQCSKLLSDTSVIQFYPSKFVLITDILDTFGKLVYI gi| 14777378 I )QKVKALKIVIQCSKLLSDTSVIQFYPSKFVLITDILDTFGKLVYI gi I 12847855 | gi|15292597| gi I 72990811 0

0

460 470 480 490 500

NOV8 gi| 12858347 | L@ESIN; HFLHTIK- gi| 14777378 | LPSETIN: FLJJTJK- gi I 12847855 I gι|15292597| SFRFVARFNfflHP gi I 72990811 jIYjPQVRPRNIKRLITFTTNLILLJ FRFVABFNHHP

510 520 530 540 550 . . | . . . . | . .

NOV8 iSHCP gi| 1285834 | QGfflELLl gi|14777378| §GHE P| ^Ξ«H^FQS!^sγi^A PraCALSΞlPSLLSgfflRGLSHl R ^DSS^Fθ3ϊSYfflA PpaAL TEij E CKlHGNNALlL gi|12847855| gi|15292597| aFD8M La|-V|ϊ]KSl| HTQDΦL ECK<|aKNNGAiLM gι|7299081| FD> «LLS|-V|||KSSLHTQD|LNECK JKNNGA|LM

560 570 580 590 600 .. I .. ..| .. ..| ..

NOV8 gι|12858347| CAEHV gi|14777378| SVMSAFRAEFIATRSMDFIGMIKECDESGFPKHLLFRSLGLNLALADPPE g I 128478551 gi|15292597| SVLSSFNSEFIATNALEFIALINASETPGISKSQLLRSLGSCVSSCPPLQ gi I 72990811 SVLISFNSEFIATNALEFIALINASETPGISKSQLLRSLGSCVSSCPPLQ

610 620 630 640 650 .. I .. ,.| .. ..| .. .. I

NOV8 gi I 12858347 | gi 114777378 | SDRLQILNEAWKVITKLKNPQDYINCAEVWVEYTCKHFT- g I 12847855 | gi|15292597| EQRVTFLKAAFETINKLTDPNEYINCVET AVFVSQYFT gi|7299081| EQRVTFLKAAFETINKLTDPNEYINCVETWAVFVSQYFTVSFKSISKILS

660 670 680 690 700 ..I .. ..I .. • •I

NOV8 gi|12858347| gi I 14777378 I KREVNTVLADVIKHMTPDRAFEDSYPQLQLIIKKVIAHFHDF gι|12847855| g I 15292597 | IHEVNRLLGELNTRMCLGKAYEKHYSQLQNILTRIMQNYRSI gi|7299081| MMLKSFYQIHEVNRLLGELNTRMCLGKAYEKHYSQLQNILTRIMQNYRSI

710 720 730 740 750 I I I I I I I I I I

NOV8 gi I 14777378 I SVLFSVEKFLPFLDMFQKEΞVRVEVCKCIMDAFIKHQQEPTKDPVILNAL gil 12847855 I gi I 15292597 I ELLLIQPNFLPYLDLFQKESVRVEVCKNILSFYKQNΞDEYTCDAVVTNAL gi I 72990811 ELLLIQPNFLPYLDLFQKESVRVEVCKNILSFYKQNSDEYTCDAVVTNAL

760 770 780 790 800

....I |....|....|....|....|....|....| I.... I

NOV8 gi 112858347 I gi I 14777378 I LHVCKTMHDSVNALTLEDEKRMLSYLINGFIKMVSFGRDFEQQLSFYVES gi 112847855 I gi I 15292597 | MYLGKILNDSVNALSVDDERRQIAQLINVFIHKVHFGNDLEQQLSFYVEA gi I 72990811 MYLGKILNDSVNALSVDDERRQIAQLINVFIHKVHFGNDLEQQLSFYVEA

810 820 830 840 850 I I I I I .... I I I I I

N0V8 gi I 12858347 I g I 14777378 I RSMFCNLEPVLVQLIHSVNRLAMETRKVMKGNHSRKTAAFVRACVAYCFI gi I 15292597 | RGTFSNLDAVYVTLVHAACKLATRNRSKSTG FVKACIAYCFI gi I 72990811 RGTFSNLDAVYVTLVHAACKLATRNRSKSTG FVKACIAYCFI

860 870 880 890 900 I I I I I ....| I I I I

N0V8 gi I 14777378 I TIPSLAGIFTRLNLYLHSGQVALANQCLSQADAFFKAAISLVPEVPK -I gi I 15292597 | TIPSIEAVQQQMNLYLLCGQLALQHLCLGQADACFEAALQLVNELPAATV gi I 72990811 TIPSIEAVQQQMNLYLLCGQLALQHLS DACFEAALQLVNELPAATV

910 920 930 940 950

....|....|....|....|.... |....|....|....|....|....|

N0V8 gi 112858347 I gi I 14777378 I NIDGKMRPSEΞFLLEFLCNFFΞTLLIVPDHPEHGVLFLVRELLNVIQDYT gi 112847855 I gi I 15292597 | DFDGKPRSLEPFLVSYMCNILATLIVVPDSPEQGVLYFLRLLLEVVGRHK gi I 72990811 DFDGKPRSLEPFLVSYMCNILATLIVVPDSPEQGVLYFLRLLLEVVGRHK

960 970 980 990 1000 I I I I I I I I I I

N0V8 gi 112858347 I gi I 14777378 | EDNSDEKIRIYTCVLHLLSAMSQETYLYHIDKVDSNDSLYGGDSKFLAE gi 112847855 I gi I 15292597 | FKVDSSAPSIIYLHSLDMLYVQSLERFPYHIKGVVSNDDLYGHDPKFLQE gi I 72990811 FKVDSSAPSIIYLHSLDMLYVQSLERFPYHIKGVVSNDDLYGHDPKFLQE

1010 1020 1030 1040 1050

....I |.... I....|....|.... |....|.... |.... |.... I

N0V8 gi 112858347 | gi I 14777378 I NNKLCETVMAQILEHLKTLAKDEALKRQSSLGLSFFNSILAHGDLRNNKL gi 112847855 I gi I 15292597 | VNNMCAQVVDAILLQLKSLGVAQQQRSQAELALELFLRIVKYADLERETI gi I 7299081 I VNNMCAQVVDAILLQLKSLGVAQQQRSQAELALELFLRIVKYADLERETI

1060 1070 1080 1090 1100 I I I I I .... I I .... I I I

N0V8 NQLSVNLWHLA-QRHGCADTRTMVKTLEYIKKQSKQPDMTHLTELALRLP AQLAVNL LLANKAQSQLDVKTLPQTLRSVEIIYKQIKDASPIRAQTIAK

AQLAVNLWLLANKAQSQLDVKTLPQTLRSVEIIYKQIKDASPIRAQTIAK

N0V8 gι|12858347| gil 14777378 I LQTRT gι|12847855| gι| 15292597 I LLLRVRSS gι| 72990811 LLLRVRSS The Seel -related proteins bind to syntaxin family t-SNAREs with high affinity, thus controlling the interaction of syntaxins with their cognate SNARE partners. Muncl8-2 is a Seel homologue enriched in epithelial cells and forms a complex with syntaxin 3, a t-SNARE localized to the apical plasma membrane. Mutational studies implied that Muncl8-2 function in apical membrane trafficking involves aspects independent ofthe syntaxin 3 interaction (Riento et al., Muncl8-2, a functional partner of syntaxin 3, controls apical membrane trafficking in epithelial cells. J Biol Chem 275(18): 13476-83, 2000).

The Q-SNARE syntaxin 1 is a central component ofthe synaptic membrane fusion machinery. Syntaxin probably interacts with multiple proteins during synaptic vesicle exocytosis. In vitro, the tightest binding partners for syntaxin 1 are other SNAREs (synaptobrevin/VAMP and SNAP-25) and muncl8-l (also known as rbsecl/nsecl) (Matos et al., The relation of protein binding to function: what is the significance of munclδ and synaptotagmin binding to syntaxin 1, and where are the corresponding binding sites? Eur J Cell Biol 79(6):377-82, 2000).

Mintl (XI 1 /human Lin- 10) and Mint2 are neuronal adaptor proteins that bind to Muncl8-1 (n/rb-secl), a protein essential for synaptic vesicle exocytosis. Mintl has previously been characterized in a complex with CASK, another adaptor protein that in turn interacts with neurexins. Neurexins are neuron-specific cell surface proteins that act as receptors for the excitatory neurotoxin -latrotoxin. Hence, one possible function for Mintl is to mediate the recruitment of Munclδ to neurexins. In agreement with this hypothesis, it was shown that the cytoplasmic tail of neurexins captures Munclδ via a multiprotein complex that involves Mintl. Furthermore, it was demonstrated that both Mintl and Mint2 can directly bind to neurexins in a PDZ-domain mediated interaction. Various Mint and/or CASK containing complexes can be assembled on neurexins, and we demonstrate that Mintl can bind to

Muncl8 and CASK simultaneously. These data support a model whereby one ofthe functions of Mints is to localize the vesicle fusion protein Munclδ to those sites at the plasma membrane that are defined by neurexins, presumably in the vicinity of points of exocytosis (Biederer and Sudhof, Mints as adaptors. Direct binding to neurexins and recruitment of muncl8. J Biol Chem 275(51):39803-6, 2000).

The above defined information for NOV8 suggests that NOV8 may function as a member of a Munclδ protein family. Therefore, the NOV8 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV8 compositions of the present invention will have efficacy for treatment of patients suffering from inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; blood disorders; asthma; psoriasis; inflammatory skin disordersvascular disorders, hypertension, skin disorders, renal disorders including Alport syndrome, immunological disorders, tissue injury, cancers, fibrosis disorders, bone diseases, Ehlers-Danlos syndrome type VI, VII, type IV, S-linked cutis laxa and Ehlers- Danlos syndrome type V, osteogenesis imperfecta, Immuno therapy of inflammatory and infectious diseases such as AIDS, treatment of Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), anorexia, bulimia, asthma, Parkinson's disease, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, multiple sclerosis; and Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, asthma, allergies, benign prostatic hypertrophy, and psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease or Gilles de la Tourette syndrome. The NOV8 nucleic acid encoding Muncl 8-like protein, and the Muncl 8-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV9

NOV9 includes two novel Immunoglobulin-like proteins disclosed below. The disclosed proteins have been named NOV9a and NOV9b.

NOV9a

A disclosed NOV9a nucleic acid of 1514 nucleotides (also referred to SC138673511_A) encoding a novel Immunoglobulin-like protein is shown in Table 9A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 29- 31 and ending with a TGA codon at nucleotides 1319-1321. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon is underlined in Table 9A, and the start and stop codons are in bold letters. Table 9A. NOV9a Nucleotide Sequence (SEQ ID NO:39)

CTCTGTAAGGGCCAGAAGTGAGGCAGGGATGGGGGTGTTGGATGCCTGGATGGGCCCAGAGGTCTGGGTCCCCCAGGGGG CAGGGGCCAAAGGTCCAGACCCCCCAAGTCCAGTGAGGGCAGTAGGGATTGGGTTGGGGGAAGATAACTGGGGGAAGGCC AGGGCCCGGCGCTTCCAGTGGCCGCTGCTGCTGCTGTGGGCGGCCGCGGCGGGGCCAGGGGCAGGACAGGAAGTACAGAC AGAGAACGTGACAGTGGCTGAGGGTGGGGTGGCTGAGATCACCTGCCGTCTGCACCAGTATGATGGGTCCATAGTTGTCA TCCAGAACCCAGCCCGGCAGACCCTCTTCTTCAATGGCACCCGTGCCTTGAAGGATGAGCGTTTCCAGCTTGAGGAGTTC TCCCCACGCCGGGTGCGGATCCGGCTCTCAGATGCCCGCCTGGAGGACGAGGGGGGCTATTTCTGCCAGCTCTACACAGA AGACACCCACCACCAGATTGCCACGCTCACGGTACTAGTGGCCCCAGAGAATCCTGTGGTGGAGGTCCGGGAGCAGGCGG TAGAGGGCGGCGAGGTGGAGCTCAGCTGCCTCGTTCCGCGGTCCCGTCCGGCTGCCACCCTGCGCTGGTACCGGGACCGC AAGGAGCTGAAAGGAGTGAGCAGCAGCCAGGAAAATGGCAAGGTCTGGAGCGTGGCAAGCACAGTACGGTTTCGTGTGGA CCGTAAGGACGACGGTGGTATCATCATCTGTGAGGCGCAGAACCAGGCGCTGCCCTCCGGACACAGCAAGCAGACGCAGT ACGTGCTGGATGTGCAGTACTCCCCCACGGCCCGGATTCATGCCTCCCAAGCTGTGGTGAGGGAGGGAGACACGCTGGTG TTGACGTGTGCTGTCACGGGGAACCCCAGGCCAAACCAGATCCGCTGGAACCGCGGGAATGAGTCTTTGCCGGAGAGGGC GGAGGCCGTGGGAGAGACGCTCACGCTGCCGGGTCTGGTATCCGCGGATAACGGCACCTACACTTGCGAGGCGTCCAATA AGCACGGCCATGCGAGGGCGCTCTACGTACTTGTGGTCTACGACCCTGGTGCGGTGGTAGAGGCTCAGACGTCGGTTCCC TATGCCATTGTGGGCGGCATCCTGGCGCTGCTGGTGTTTCTGATCATATGTGTGCTAGTGGGCATGGTCTGGTGCTCGGT ACGGCAGAAGGGTTCCTATCTGACCCACGAAGCCAGTGGCTTGGATGAACAGGGAGAAGCAAGAGAAGCCTTCCTCAATG GCAGCGACGGACACAAGAGGAAAGAGGAATTCTTCATCTGACCCTATCCCCAGCCAGCCTAGGCCTGGGCCTGGGCTGGG GTCCCCCCCACTGCCAGCTGCAAGGAACCAGCAAAGACATTTACCAGAGTCTGGGATGGTGGGCTTCTCCCCCCACCACT AACACCTCAGACGCTTGGGCAGGGATGGGGGTGTTGGATGCCTGGATCTCTGTAAGGGCCAGAAGTGAGGGCCC

The disclosed NOV9a nucleic acid sequence, maps to chromosome 19, has 510 of 852 bases (59%) identical to a Mus musculus immunosuperfamily protein B12 mRNA from (gb:GENBANK-ID:AF061260|acc:AF061260) (E = 7.6e^"21).

A disclosed NOV9a polypeptide (SEQ ID NO:40) encoded by SEQ ID NO:39 is 430 amino acid residues and is presented using the one-letter amino acid code in Table 9B. Signal P, Psort and/or Hydropathy results predict that NOV9a contains a signal peptide and is likely to be localized in the plasma membrane with a certainty of 0.7300. The most likely cleavage site for a NOV9a peptide is between amino acids 66 and 67, at: GAG-QE.

Table 9B. Encoded NOV9a protein sequence (SEQ ID NO:40).

MGVLDAWMGPEVWVPQGAGAKGPDPPSPVRAVGIGLGEDNWGKARARRFQWPLLLLWAAAAGPGAGQEVQTENVTVAEGG VAEITCRLHQYDGSIVVIQNPARQTLFFNGTRALKDERFQLEEFSPRRVRIRLSDARLEDEGGYFCQLYTEDTHHQIATL TVLVAPENPVVEVREQAVEGGEVELΞCLVPRSRPAATLRWYRDRKELKGVSSSQENGKVWSVASTVRFRVDRKDDGGIII CEAQNQALPSGHSKQTQYVLDVQYSPTARIHAΞQAVVREGDTLVLTCAVTGNPRPNQIRWNRGNESLPERAEAVGETLTL PGLVSADNGTYTCEASNKHGHARALYVLVVYDPGAVVEAQTSVPYAIVGGILALLVFLIICVLVGMV CSVRQKGSYLTH EASGLDEQGEAREAFLNGΞDGHKRKEEFFI

The NOV9a amino acid sequence has 366 of 381 amino acid residues (96%) identical to, and 366 of 381 amino acid residues (96%) similar to, the Homo sapiens 381 amino acid residue F22162 protein (ptnr:SPTREMBL-ACC:Q9Y4A4) (E = 2.8e^'191).

NOV9a is expressed in at least the following tissues: Amygdala, Brain, Coronary Artery, Heart, Hippocampus, Hypothalamus, Kidney, Lung, Pituitary Gland, Spinal Chord, Substantia Nigra, Thalamus, Whole Organism, SeqCalling_celltypes: glioblastoma total_fetus schizo brain brain oligodendroglioma Fetal brain. This information was derived by determining the tissue sources ofthe sequences that were included in the invention. SeqCalling sources: Amygdala, Brain, Coronary Artery, Heart, Hippocampus, Hypothalamus, Kidney, Lung, Pituitary Gland, Spinal Chord, Substantia Nigra, Thalamus, Whole Organism, PublicEST sources: glioblastoma total_fetus schizo brain brain oligodendroglioma Fetal brain. In addition, NOV9a is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus immunosuperfamily protein B12 mRNA (GENBANK-ID: gb:GENBANK-ID:AF061260|acc:AF061260): glioblastoma total_fetus schizo brain brain oligodendroglioma Fetal brain.

NOV9b

A disclosed NOV9b nucleic acid of 1161 nucleotides (also referred to CG106625-02) encoding a novel Immunoglobulin-like protein is shown in Table 9C. An open reading frame lacking the signal peptide was identified beginning with an GCC codon at nucleotides 2-4 and ending with a TGA codon at nucleotides 1157-1159. Putative untranslated regions upstream from the initiation codon and downstream from the termination codon is underlined in Table 9C, and the start and stop codons are in bold letters.

Table 9C. NOV9b Nucleotide Sequence (SEQ ID NO:41)

GGCCCGGCGCTTCCAGTGGCCGCTGCTGCTGCTGTGGGCGGCCGCGGCGGGGCCAGGGGCAGGACAGGAAGTACAGACAG AGAACGTGACAGTGGCTGAGGGTGGGGTGGCTGAGATCACCTGCCGTCTGCACCAGTATGATGGGTCCATAGTTGTCATC CAGAACCCAGCCCGGCAGACCCTCTTCTTCAATGGCACCCGTGCCTTGAAGGATGAGCGTTTCCAGCTTGAGGAGTTCTC CCCACGCCGGGTGCGGATCCGGCTCTCAGATGCCCGCCTGGAGGACGAGGGGGGCTATTTCTGCCAGCTCTACACAGAAG ACACCCACCACCAGATTGCCACGCTCACGGTACTAGTGGCCCCAGAGAATCCTGTGGTGGAGGTCCGGGAGCAGGCGGTA GAGGGCGGCGAGGTGGAGCTCAGCTGCCTCGTTCCGCGGTCCCGTCCGGCTGCCACCCTGCGCTGGTACCGGGACCGCAA GGAGCTGAAAGGAGTGAGCAGCAGCCAGGAAAATGGCAAGGTCTGGAGCGTGGCAAGCACAGTACGGTTTCGTGTGGACC GTAAGGACGACGGTGGTATCATCATCTGTGAGGCGCAGAACCAGGCGCTGCCCTCCGGACACAGCAAGCAGACGCAGTAC GTGCTGGATGTGCAGTACTCCCCCACGGCCCGGATTCATGCCTCCCAAGCTGTGGTGAGGGAGGGAGACACGCTGGTGTT GACGTGTGCTGTCACGGGGAACCCCAGGCCAAACCAGATCCGCTGGAACCGCGGGAATGAGTCTTTGCCGGAGAGGGCGG AGGCCGTGGGAGAGACGCTCACGCTGCCGGGTCTGGTATCCGCGGATAACGGCACCTACACTTGCGAGGCGTCCAATAAG CACGGCCATGCGAGGGCGCTCTACGTACTTGTGGTCTACGACCCTGGTGCGGTGGTAGAGGCTCAGACGTCGGTTCCCTA TGCCATTGTGGGCGGCATCCTGGCGCTGCTGGTGTTTCTGATCATATGTGTGCTAGTGGGCATGGTCTGGTGCTCGGTAC GGCAGAAGGGTTCCTATCTGACCCACGAAGCCAGTGGCTTGGATGAACAGGGAGAAGCAAGAGAAGCCTTCCTCAATGGC AGCGACGGACACAAGAGGAAAGAGGAATTCTTCATCTGACC

The disclosed NOV9b nucleic acid sequence, maps to chromosome 19, has 557 of 931 bases (59%) identical to a Mus musculus sgigsf mRNA for spermatogenic immunoglobulin superfamily protein (gb:GENBANK-ID:AB052293|acc:AB052293.1) (E = 2.1e^"26).

A disclosed NOV9b polypeptide (SEQ ID NO:42) encoded by SEQ ID NO:41 is 385 amino acid residues and is presented using the one-letter amino acid code in Table 9D. Signal P, Psort and/or Hydropathy results predict that NOV9b contains a signal peptide and is likely to be localized in the plasma membrane with a certainty of 0.4600. The most likely cleavage site for a NOV9b peptide is between amino acids 21 and 22, at: GAG-QE.

Table 9D. Encoded NOV9b protein sequence (SEQ ID NO:42).

ARRFQ PLLLL AAAAGPGAGQEVQTENVTVAEGGVAEITCRLHQYDGSIVVIQNPARQTLFFNGTRALKDERFQLEEFS PRRVRIRLSDARLEDEGGYFCQLYTEDTHHQIATLTVLVAPENPVVEVREQAVEGGEVELSCLVPRSRPAATLRWYRDRK ELKGVSSSQENGKV SVASTVRFRVDRKDDGGIIICEAQNQALPSGHSKQTQYVLDVQYSPTARIHASQAVVREGDTLVL TCAVTGNPRPNQIR NRGNESLPERAEAVGETLTLPGLVSADNGTYTCEASNKHGHARALYVLVVYDPGAVVEAQTSVPY AIVGGILALLVFLIICVLVGMVWCSVRQKGSYLTHEASGLDEQGEAREAFLNGSDGHKRKEEFFI The NOV9b amino acid sequence has 366 of 381 amino acid residues (96%) identical to, and 366 of 381 amino acid residues (96%) similar to, the Homo sapiens 381 amino acid residue F22162_l protein (ptnr:SPTREMBL-ACC:Q9Y4A4) (E = 3.7e^"191).

NOV9b is expressed in at least the following tissues: Amygdala, Brain, Coronary Artery, Heart, Hippocampus, Hypothalamus, Kidney, Lung, Pituitary Gland, Spinal Chord, Substantia Nigra, Thalamus, Whole Organism. This information was derived by determining the tissue sources ofthe sequences that were included in the invention. SeqCalling sources: Amygdala, Brain, Coronary Artery, Heart, Hippocampus, Hypothalamus, Kidney, Lung, Pituitary Gland, Spinal Chord, Substantia Nigra, Thalamus, Whole Organism, PublicEST sources: glioblastoma total_fetus schizo brain brain oligodendroglioma Fetal brain. In addition, NOV9b is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus sgigsf mRNA for spermatogenic immunoglobulin superfamily protein (gb:GENBANK-ID:AB052293|acc:AB052293.1): Fetal brain.

NOV9a and NOV9b are very closely homologous as is shown in the amino acid alignment in Table 9E.

Table 9E Amino Acid Alignment of NOV9a and NOV9b

10 20 30 40 50 I I I I I I .... I I I • I

NOV9a GVLDA MGPEVWVPQGAGAKGPDPPSPVRAVGIGLGEDN GKAR NOV9b

110 120 130 140 150

210 220 230 240 250

310 320 330 340 350 ..I..

NOV9a NRGNESLPERAEAVGETLTLPGLVSADNGTYTCEAΞNKHGHARALYVLV\j NOV9b NRGNESLPERAEAVGETLTLPGLVSADNGTYTCEASNKHGHARALYVLVV

360 370 380 390 400

410 420 430 .. I .. .. |.. I

NOV9a iSGLDEQGEAREAFLNGSDGHKRK NOV9b SGLDEOGEAREAFLNGSDGHKRKI

Homologies to any ofthe above NOV9 proteins will be shared by the other NOV9 proteins insofar as they are homologous to each other as shown above. Any reference to NOV9 is assumed to refer to both ofthe NOV9 proteins in general, unless otherwise noted.

NOV9a also has homology to the amino acid sequences shown in the BLASTP data listed in Table 9F.

The homology of these sequences is shown graphically in the ClustalW analysis shown in Table 9G.

Table 9G Information for the ClustalW proteins

1) NOV9a (SEQ ID NO.40)

2) gιl3451335lgblAAC32740.1l (AC005525) F22162 [Homo sapiens] (SEQ ID NO:109)

3) gιl7767239lgb!AAF69029.1 IAF 132811 1 (AF 132811) nectin-hke protein 2 [Homo sapiens] (SEQ ID NO.110)

4) gιl7657226|ref|NP 055148.1| (NM_014333) immunoglobuhn superfamily, member 4 [Homo sapiens] (SEQ ID NO 111)

5) gιll4328885[dbιlBAB60686 II (AB052293) spermatogenic immunoglobuhn superfamily protem [Mus musculus] (SEQ ID NO: 112)

6) gιll2851464ldbιlBAB290501l (AK013911) putative [Mus musculus] (SEQ ID NO.113)

10 20 30 40 50

...|....|....|....|....|....|....|....|....|

NOV9a -MGVLDAWMGPEVW§jPQ§i§G AKGPDPPSPVR0VGIG-- gι|3451335| gι|7767239| ASVvlPSffls QCA gι|7657226| MASVVfiPStg QCA gι|14328885| — MASAViPSeS QCA W— gi I 12851464 | CHDWSLLTPPPAHLISISLAgRSg|GGSHRRQSEAGARHGECC0AERIPV

gi I 14328885 I AAAAAPPGL|L|LLΠ gι|12851464| CGGTAVAAAAAPPGLgLgLLy

160 170 180 190 200

210 220 230 240 250

310 320 330 340 350

360 370 380 390 400

Tables 9H - 9P list the domain description from DOMAIN analysis results against NOV9. This indicates that the NOV9 sequence has properties similar to those of other proteins known to contain these domains.

Table 9H. Domain Analysis of NOV9a qnl I Smart |smart00408, IGc2, Immunoglobulin C-2 Type (SEQ ID NO: 114)

Length = 63 residues, 100.0% aligned

Score = 63.2 bits (152), Expect = 3e-ll

NOV9a 278 REGDTLVLTCAVTGNPRPNQIRWNRGNESLPE-RAEAVGETLTLPGLVSADNGTYTCEAS 336

I I +++ I I I +1 + 1 II I I + + 111 I I I 111+ + l + l III I 00408 1 LEGESVTLTCPASGDPVPN-IT LKDGKPLPESRVVASGSTLTIKNVSLEDSGLYTCVAR 59

NOV9a 337 NKHG 340

I I 00408 60 NSVG 63

Table 91. Domain Analysis of NOV9a gnl I Smart I smart00408, IGc2, Immunoglobulin C-2 Type (SEQ ID NO: 115)

Length = 63 residues, 96.8% aligned

Score = 40.4 bits (93), Expect = 2e-04

NOV9a 179 EGGEVELSCLVPRSRPAATLRWYRDRKELKGVSSSQENGKVWSVASTVRFRVDRKDDGGI 238

I I I l + l I + I +1 I I I 1+ + +1 1 +

00408 2 EGESVTLTCPA-SGDPVPNIT LKDG PLPESRVVASG STLTIKNVSLEDSGL 53

NOV9a 239 IICEAQNQA 247

I l + l 00408 54 YTCVARNSV 62

Table 9J. Domain Analysis of NOV9a gnl I Smart I smart00409, IG, Immunoglobulin (SEQ ID NO: 116)

Length = 86 residues, 98.8% aligned

Score = 60.5 bits (145), Expect = 2e-10

NOV9a 273 SQAVVREGDTLVLTCAVTGNPRPNQIR NRGNESL PERAEAVGE TLTLPGLVS 325 l + l I+++ l + l +1 I I I + I + I I I I 1+ +

00409 2 PSVTVKEGESVTLSCEASGNPPP-TVT YKQGGKLLAESGRFSVSRSGGNSTLTISNVTP 60

NOV9a 326 ADNGTYTCEASNKHGHARALYVLVVY 351 l + l I I I I l + l 1 1 + I I 00409 61 EDSGTYTCAATNSSGSASSGTTLTVL 86

Table 9K. Domain Analysis of NOV9a gnl I Smart | smart00409, IG, Immunoglobulin (SEQ ID NO: 117)

Length = 86 residues, 98.8% aligned

Score = 44.7 bits (104), Expect = le-05

NOV9a 72 ENVTVAEGGVAEITCRLHQYDGSIVVIQNPARQTLFFNGTRALKDERFQLEEFSPRRVRI 131

+III II ++I I + 1 +1 M + l + 00409 2 PSVTVKEGESVTLSCEASGNPPPTVTWYKQGGKLLAESG RFSVSR-SGGNSTL 53

NOV9a 132 RLSDARLEDEGGYFCQLYTED-THHQIATLTVL 163

+1+ ll l l l + ll lll 00409 54 TISNVTPEDSGTYTCAATNSSGSASSGTTLTVL 86 Table 9L. Domain Analysis of NOV9a gnl I Smart I smart00409, IG, Immunoglobulin (SEQ ID NO: 118)

Length = 86 residues, 77.9% aligned

Score = 36.6 bits (83), Expect = 0.003

NOV9a 177 AVEGGEVELSCLVPRSRPAATLRWYRDRKELKGVSSSQENGKV SVASTVRFRVD--RKD 234

I I I I I I I 1+ I 1+ +1 +^• I + +

00409 6 VKEGESVTLSCEAS-GNPPPTVTWYKQGGKL LAESGRFSVSRSGGNSTLTISNVTPE 61

NOV9a 235 DGGIIICEAQN 245

I I I I I 00409 62 DSGTYTCAATN 72

Table 9M. Domain Analysis of NOV9a gnl I Pfam|pfam00047, ig, Immunoglobulin domain. Members of the immunoglobulin' superfamily are found in hundreds of proteins of different functions. Examples include antibodies, the giant muscle kinase titin and receptor tyrosine kinases. Immunoglobulin-like domains may be involved in protein-protein and protein-ligand interactions. The Pfam alignments do not include the first and last strand of the immunoglobulin-like domain. (SEQ ID NO: 119) Length = 68 residues, 100.0% aligned Score = 38.9 bits (89), Expect = 6e-04

NOV9a 180 GGEVELSCLVPRSRPAATLR YRDRKELKGVSSSQE NGKVWSVASTVRFRVDRKDD 235

I 1 1+1 1 I I+ I II II++ + II+ 1+ + ++ +1 00047 1 GESVTLTCSVSGYPPDPTVTWLRDGKEIELLGSSESRVSSGGRFSISSLSLTISSVTPED 60

NOV9a 236 GGIIICEA 243

I I 00047 61 SGTYTCVV 68

Table 9N. Domain Analysis of NOV9a gnl I Pfam|pfam00047, ig, Immunoglobulin domain. (SEQ ID NO: 120)

Length = 68 residues, 100.0% aligned

Score = 38.9 bits (89), Expect = 6e-04

NOV9a 280 GDTLVLTCAVTGNPRPNQIRWNRGNE SLPERAEAVGETLTLPGLVSAD 327

I+++ I I l + l + l I + 1 1 + I I +11+ + I

00047 1 GESVTLTCSVSGYPPDPTVTWLRDGKEIELLGSSESRVSSGGRFSISSLSLTISSVTPED 60

NOV9a 328 NGTYTCEA 335

+ 1 I I I I 00047 61 SGTYTCVV 68

Table 9O. Domain Analysis of NOV9a gnl I Smart | smart00294, 4.1m, putative band 4.1 homologues' binding motif (SEQ ID NO: 121)

Length = 19 residues, 100.0% aligned

Score = 38.5 bits (88), Expect = 8e-04

NOV9a 386 MVWCSVRQKGSYLTHEASG 404

I I +1 I I I II I 00294 1 MYRYKHRDEGSYHTHEPKG 19 Table 9P. Domain Analysis of NOV9a gnl I Smart I smart00406, IGv, Immunoglobulin V-Type (SEQ ID NO: 122)

Length = 80 residues, 97.5% aligned

Score = 37.0 bits (84), Expect = 0.002

NOV9a 82 AEITCRLHQYDGSIVVI QNPARQTLFF NGTRALKDERFQL-EEFSPRRV 129

++I+ + | + 1 1 + + + + I I + ++ I I 00406 2 VTLSCKASGFTFSSYYVS VRQPPGKGLE LGYIGSDVSYSEASYKGRVTISKDNSKNDV 61

NOV9a 130 RIRLSDARLEDEGGYFCQ 147

+ +1+ l+l I I l+l 00406 62 SLTISNLRVEDTGTYYCA 79

The effect of rIL-4 on the expression of low affinity receptor for the Fc part of IgE (Fc epsilon R2/CD23) and class II MHC antigens on Burkitt's lymphoma (BL) cell lines was investigated. Some ofthe BL lines contained low percentages of CD23 and HLA-DQ-positive cells, but virtually all cells expressed HLA-DR. IL-4 induced CD23 and class II MHC Ag expression on 7 of 9 BL. Optimal CD23 and class II MHC expression was observed after 48- 72 h of incubation. Induction of CD23 and class II MHC Ag in the BL cell line BL2 by IL-4 was confirmed at the specific mRNA level. Significant activation of HLA-DQ mRNA was obtained after 6 h of incubation with IL-4 and gradually increased during prolonged incubation. Maximal induction of mRNA transcription occurred after 48 to 72 h. Optimal induction of HLA-DR and CD23 transcription in BL2 was also observed after 48 to 72 h. The induction of CD23 and class II MHC Ag seems to be specific for IL-4, because rIL-1, rIL-2, rIFN-gamma, recombinant granulocyte-macrophage-CSF, and a commercial source of low m.w. B cell growth factor were ineffective. In addition, the expression of class I MHC Ag, the transferrin receptor, CD38, CD25, CD10, CD20, and CD21 were not affected by IL-4. Interestingly, IFN-gamma and PGE2 suppressed the IL-4-induced membrane expression of CD23 and class II MHC Ag in a dose-dependent way. IFN-gamma also blocked IL-4-induced CD23 mRNA transcription in BL2 completely, whereas PGE2 (10(-7) M) was partially inhibitory. The induction of CD23 and class II MHC Ag by IL-4 required intact protein synthesis as shown by its inhibition by cycloheximide. These results indicate that the induction of CD23 and class II MHC Ag by IL-4 is regulated in a coordinated way (Rousset et al., Regulation of Fc receptor for IgE (CD23) and class II MHC antigen expression on Burkitt's lymphoma cell lines by human IL-4 and IFN-gamma. J Immunol 140(8):2625-32, 1988). The B cell surface trigger(s) and the molecular mechanism(s) of somatic hypermutation remain unknown, partly because ofthe lack of amendable in vitro models. Recently, however, it was reported that upon B cell receptor cross-linking and coculture with activated T cells, the Burkitt's lymphoma cell line BL2 introduces mutations in its IgVH gene in vitro. The relevance of a culture model is confirmed by establishing that the entire spectrum of somatic mutations observed in vivo, including insertions and deletions, could be found in the DNA of BL2 cells. Additionally, it was showed that among four human B cell lines, only two with a centroblast-like phenotype can be induced to mutate. Triggering of somatic mutations in BL2 cells requires intimate T-B cell contacts and is independent of CD40-CD40- ligand (CD40L) interactions as shown by 1) the lack of effect of anti-CD40 and/or anti-

CD40L blocking Abs on somatic mutation and 2) the ability of a CD40L-defιcient T cell clone (isolated from an X-linked hyper-IgM syndrome patient) to induce somatic mutation in B cell receptor-engaged BL2 cells. Thus, the in vitro model reveals that T-B cell membrane interactions through surface molecules different from CD40-CD40L can trigger somatic hypermutation (Denepoux et al., T cells can induce somatic mutation in B cell receptor- engaged BL2 Burkitt's lymphoma cells independently of CD40-CD40 ligand interactions. J Immunol 164(3):1306-13, 2000).

A hybridoma-derived monoclonal antibody, produced by immunization with the Burkitt's tumor-derived B-lymphoblastoid cell line, B35M, was previously shown to detect a 68,000 dalton surface membrane protein, BL2, on the surface of peripheral blood B cells, which is absent from thymocytes, T cells, and granulocytes. In a recent study, the expression and distribution of BL2 on benign and malignant human lymphoid cells was investigated. Indirect immunofluorescent assay with this monoclonal antibody demonstrated that BL2 is expressed by cells within the fetal liver and by a variable proportion of lymph node, tonsil, and spleen B cells, but not by T cells. The neoplastic cells isolated from 18 T-cell malignancies were BL2- . BL2 was was hetero geneously expressed by a variable proportion ofthe malignant cells in 29/32 cases of B-chronic lymphocytic leukemia and 33/38 cases of B-cell lymphomas, but appeared to be lost in the terminal stages of B-cell differentiation, as myeloma plasma cells were BL2- . BL2 expression was not limited to B cells of a particular surface immunoglobulin isotype. Immunofluorescent staining for BL2 in cryostat tissue sections demonstrated that the majority, but not all, germinal center and interfollicular Ia+ (non-T) cells are BL2+. These findings suggested that BL2 is a B-cell lineage-specific differentiation marker that may be useful in the study of B-cell ontogeny and in defining subgroups ofthe B-cell malignancies (Knowles et al., A new human B-lymphocyte surface antigen (BL 2) detectable by a hybridoma monoclonal antibody: distribution on benign and malignant lymphoid cells. Blood 1983 Jul;62(l):191-9, 1983).

The protein similarity information, expression pattern, and map location for the NOV9 suggest that NOV9 may have important structural and/or physiological functions characteristic ofthe Immunoglobulin protein family. Therefore, the NOV9 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV9 compositions of the present invention will have efficacy for treatment of patients suffering from inflammatory disorders such as osteo- and rheumatoid-arthritis, inflammatory bowel disease, Crohn's disease; immunological disorders, AIDS; cancers including but not limited to lung cancer, colon cancer, leukemia or pancreatic cancer.; blood disorders; asthma; psoriasis; inflammatory skin disordersvascular disorders, hypertension, skin disorders, renal disorders including Alport syndrome, immunological disorders, tissue injury, cancers, fibrosis disorders, bone diseases, Ehlers-Danlos syndrome type VI, VII, type IV, S-linked cutis laxa and Ehlers- Danlos syndrome type V, osteogenesis imperfecta, Immuno therapy of inflammatory and infectious diseases such as AIDS, treatment of Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), anorexia, bulimia, asthma, Parkinson's disease, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, multiple sclerosis; and Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, asthma, allergies, benign prostatic hypertrophy, and psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease or Gilles de la Tourette syndrome. The NOV9 nucleic acid encoding Immunoglobulin-like protein, and the Immunoglobulin-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOV10

NOV 10 includes three novel Type II Cytokeratin-like proteins disclosed below. The disclosed proteins have been named NOV 10a, NOV 10b and NOV 10c.

NOVlOa A disclosed NOV 10a nucleic acid of 1782 nucleotides (also referred to

GSAC055715.12_D) encoding a Type II Cytokeratin-like protein is shown in Table 10A. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 63- 65 and ending with a TAA codon at nucleotides 1710-1712. Putative untranslated regions upstream from the intiation codon and downstream from the termination codon are underlined in Table IOA, and the start and stop codons are in bold letters.

Table IOA. NOVlOa Nucleotide Sequence (SEQ ID NO:43)

CAGAACCTTTTGGGATTTTGCCTTCCTCCCTCCCGCATCTGAGCTTTGTCTCCACCAGCAACATGAGCCGCCAATTCACC TGCAAGTCGGGAGCTGCCGCCAAGGGGGGCTTCAGTGGCTGCTCAGCTGTGCTCTCAGGGGGCAGCTCATCCTCCTTCCG GGCAGGGAGCAAAGGGCTCAGTGGGGGCTTTGGCAGCCGGAGCCTCGCAGGGAGCAAAGGGCTCAGTGGGGGCTTTGGCA GCCGGAGCCTCTACAGCCTGGGGGGTGTCCGGAGCCTCAATGTGGCCAGTGGCAGCGGGAAGAGTGGAGGCTATGGATTT GGCCGGGGCCGGGCCAGTGGCTTTGCTGGAAGCATGTTTGGCAGTGTGGCCCTGGGGCCTGTGTGCCCAACTGTATGCCC ACCTGGAGGCATCCACCAGGTTACCATCAATGAGAGCCTCCTGGCCCCCCTCAACGTGGAGCTGGACCCCAAGATCCAGA AAGTGCGTGCCCAGGAGCGAGAGCAGATCAAGGCTCTGAACAACAAGTTCGCCTCCTTCATCGACAAGGTGCGGTTCCTG GAGCAGCAGAACCAGGTACTGGAGACCAAGTGGGAGCTGCTGCAGCAGCTGGACCTGAACAACTGCAAGAACAACCTGGA GCCCATCCTCGAGGGCTACATCAGCAACCTGCGGAAGCAGCTGGAGACGCTGTCTGGGGACAGGGTGAGGCTGGACTCGG AGCTGAGGAATGTGCGGGACGTAGTGGAGGACTACAAGAAGAGGTATGAGGAGGAAATCAACAAGCGGACAGCAGCAGAG AACGAGTTTGTGCTGCTCAAGAAGGATGTGGATGCTGCTTACGCCAATAAGGTGGAACTGCAGGCCAAGGTGGAATCCAT GGACCAGGAGATCAAGTTCTTCAGGTGTCTCTTTGAAGCCGAGATCACTCAGATCCAGTCCCACATCAGTGACATGTCTG TCATCCTGTCCATGGACAACAACCGGAACCTAGACCTGGACAGCATCATTGACGAAGTCCGCACCCAGTATGAGGAGATT GCCTTGAAGAGTAAGGCCGAGGCTGAGGCCCTGTACCAGACCAAGTTCCAAGAGCTTCAGCTGGCAGCTGGCAGGCATGG GGACGACCTCAAAAACACCAAGAATGAAATCTCGGAGCTCACTCGGCTCATCCAGAGAATCCGCTCAGAGATCGAGAACG TGAAGAAGCAGGCTTCCAACCTGGAGACAGCCATCGCTGATGCTGAGCAGCGGGGAGACAACGCCCTGAAGGATGCCCGG GCCAAGCTGGACGAGCTGGAGGGCGCCCTGCACCAGGCCAAGGAGGAGCTGGCACGGATGCTGCGCGAGTACCAGGAGCT CATGAGCCTGAAGCTGGCCCTGGACATGGAGATCGCCACCTATCGCAAGCTACTGGAGAGCGAGGAGTGCAGGATGTCAG GAGAATTTCCCTCCCCTGTCAGCATCTCCATCATCAGCAGCACCAGTGGCGGCAGTGTCAGTGGCTATGGCGGTGCCAGT GGTGTCGGCAGTGGCTTAGGCCTGGGTGGAGGAAGCAGCTACTCCTATGGCAGTGGTCTTGGCGTTGGAGGTGGCTTCAG TTCCAGCAGTGGCAGAGCCATTGGGGGTGGCCTCAGCTCTGTTGGAGGCGGCAGTTCCACCATCAAGTACACCACCACCT CCTCCTCCAGCAGGAAGAGCTATAAGCACTAAAGTGCGTCTGCTAGCTCTCGGTCCCACAGTCCTCAGGCCCCTCTCTGG CTGCAGAGCCCTCTCCTCAGGT

The disclosed NOVlOa nucleic acid sequence, localized to chromosome 12, has 1255 of 1478 bases (84%) identical Mus musculus type II cytokeratin mRNA (gb:GENBANK- ID:AB033744|acc:AB033744.1) (E = 5.2e^"231).

A disclosed NOVlOa polypeptide (SEQ ID NO:44) encoded by SEQ ID NO:43 is 549 amino acid residues and is presented using the one-letter amino acid code in Table 10B. Signal P, Psort and/or Hydropathy results predict that NOVlOa does not contain a signal peptide and is likely to be localized in the cytoplasm with a certainty of 0.4500.

Table 10B. Encoded NOVlOa protein sequence (SEQ ID NO:44).

MSRQFTCKSGAAAKGGFSGCSAVLSGGSSSSFRAGSKGLSGGFGSRSLAGSKGLSGGFGSRSLYSLGGVRSLNVASGSGK SGGYGFGRGRASGFAGS FGSVALGPVCPTVCPPGGIHQVTINESLLAPLNVELDPKIQKVRAQEREQIKALNNKFASFI DKVRFLEQQNQVLETK ELLQQLDLNNCKNNLEPILEGYISNLRKQLETLSGDRVRLDSELRNVRDVVEDYKKRYEEEIN KRTAAENEFVLLKKDVDAAYANKVELQAKVESMDQEIKFFRCLFEAEITQIQSHISDMSVILS DNNRNLDLDSIIDEVR TQYEEIALKΞKAEAEALYQTKFQELQLAAGRHGDDLKNTKNEISELTRLIQRIRSEIENVKKQASNLETAIADAEQRGDN ALKDARAKLDELEGALHQAKEELARMLREYQELMSLKLALD EIATYRKLLESEECRMSGEFPSPVSISIISSTSGGSVS GYGGASGVGSGLGLGGGSΞYSYGSGLGVGGGFSSSSGRAIGGGLSSVGGGSSTIKYTTTSSSSRKSYKH

The NOVlOa amino acid sequence has 440 of 519 amino acid residues (84%) identical to, and 465 of 519 amino acid residues (89%) similar to, a Mus musculus 524 amino acid residue Type II cytokeratin (ptnr:SPTREMBL-ACC:Q9R0H5) (E = 4.6e^"221).

NOVlOa is expressed in at least the following tissues: skin, muscle, bone, cartilage, Colon carcinoma, lung. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Genomic Clone sources, Literature sources, and/or RACE sources. In addition, NOVlOa is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus mRNA for type II cytokeratin (GENBANK-ID: gb:GENBANK-ID:AB033744 |acc:AB033744.1):skin, muscle, bone, cartilage, Colon carcinoma, lung.

NOVlOb

A disclosed NOV 10b nucleic acid of 1601 nucleotides (also referred to GSAC055715 C) encoding a Type II Cytokeratin-like protein is shown in Table IOC. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 66- 68 and ending with a TGA codon at nucleotides 1599-1601. A putative untranslated region upstream from the intiation codon is underlined in Table 10C, and the start and stop codons are in bold letters.

Table IOC. NOVlOb Nucleotide Sequence (SEQ ID NO:45)

CAGGAGGGATTCATGGGGAGTTCACGGTGGAACAGACAAAAATCTCGCACCGGTACTTGCGAGCCATGAGCCGCCAACTG ACCCATTTCCCCCGCGGGGAGCGCCTGGGCTTCAGCGGTTGCTCCGCGGTCCTCTCTGGCGGGATCGGCAGCAGCTCCGC CTCATTCCGGGCCCGGGTCAAGGGCTCGGCCTCCTTTGGCAGCAAGAGCCTCTCCTGCCTTGGGGGCAGCCGAAGCCTGG CGCTCAGCGCTGCTGCACGGCGGGGCGGCGGCCGCCTGGGCGGCTTCGTGGGCACCGCCTTCGGCAGCGCCGGGCTGGGG CCCAAGTGTCCCTCCGTGTGCCCACCCGGGGGCATCCCTCAGGTCACCGTCAACAAGAGCCTCCTGGCCCCGCTCAACGT GGAGATGGACCCCGAGATCCAGAGGGTGCGCGCCCAGGAGCGGGAGCAGATCAAGGCGCTAAACAACAAGTTCGCCTCCT TCATCGACAAGGTGCGGTTCCTGGAGCAGCAGAATCAGGTGCTAGAGACCAAGTGGAACCTCCTACAGCAGCTGGACTTG AACAACTGCAGGAAGAACCTGGAGCCCATTTATGAGGGCTACATCAGCAACCTGCAGAAGCAGCTGGAGATGCTGTCTGG GGACGGGGTGAGGCTGGATTCGGAGCTGAGGAACATGCAGGATTTGGTGGAGGACTACAAGAAGAGATATGAGGTGGAGA TTAACAGACGCACAGCTGCTGAGAATGAGTTTGTGGTGCTCAAGAAGGACGTGGATGCTGCTTACATGAATAAGGTTGAG CTCCAGGCCAAGGTGGACTCCTTGACAGATGAGATTAAATTCTTCAAGTGCCTTTATGAAGGGGAGATCACTCAGATCCA GTCCCACATCAGCGACACGTCCATCGTCCTGTCAATGGACAACAACCGGGATCTGGACCTGGACAGCATCATTGCCGAGG TCCGTGCCCAGTACGAGGAGATTGCCCTAAAGAGCAAGGCCGAGGCTGAGACCCTGTACCAGACCAAGATCCAGGAGCTG CAGGTCACAGCAGGCCAGCATGGGGATGACCTCAAGCTCACCAAGGCTGAAATCTCTGAGCTCAACCGCCTGATCCAGAG GATCCGCTCAGAGATAGGGAATGTGAAGAAGCAGTGTGCCGATCTGGAGACGGCCATCGCCGACGCTGAACAGCGGGGGG ACTGCGCCCTGAAAGATGCCCGGGCCAAGCTGGATGAGCTGGAGGGCGCCCTGCACCAGGCCAAGGAGGAGCTGGCACGG ATGCTGCGTGAGTACCAGGAGCTCGTGAGCCTGAAGCTGGCCCTGGATATGGAGATCGCCACCTACCGCAAGCTGCTGGA GAGCGAGGAGTGCAGGATGTCTGGCGAATATCCAAATTCTGTGAGCATCTCCGTCATCAGCAGCACCAATGCTGGGGCAG GAGGGGCTGGCTTCAGCATGGGCTTTGGCGCCTCAAGCAGTTATAGCTACAAAACTGCAGCTGCAGACGTCAAGACCAAA GGCAGCTGTGGCAGTGAGCTCAAGGATCCCCTTGCCAAAACCTCGGGGAGCAGCTGTGCCACCAAAAAGGCCTCCAGATG A

The disclosed NOVlOb nucleic acid sequence, localized to chromosome 12, has 1004 of 1224 bases (82%) identical Mus musculus type II cytokeratin mRNA (gb:GENBANK- ID:AB033744|acc:AB033744.1) (E = 4.8e^"176). A disclosed NOVlOb polypeptide (SEQ ID NO:46) encoded by SEQ ID NO:45 is 511 amino acid residues and is presented using the one-letter amino acid code in Table 10D. Signal P, Psort and/or Hydropathy results predict that NOVlOb does not contain a signal peptide and is likely to be localized in the cytoplasm with a certainty of 0.4500.

Table 10D. Encoded NOVlOb protein sequence (SEQ ID NO:46).

MSRQLTHFPRGERLGFSGCSAVLSGGIGSSSASFRARVKGSASFGSKSLSCLGGSRSLALSAAARRGGGRLGGFVGTAFG SAGLGPKCPSVCPPGGIPQVTVNKSLLAPLNVEMDPEIQRVRAQEREQIKALNNKFASFIDKVRFLEQQNQVLETK NLL QQLDLNNCRKNLEPIYEGYISNLQKQLEMLSGDGVRLDSELRNMQDLVEDYKKRYEVEINRRTAAENEFVVLKKDVDAAY MNKVELQAKVDSLTDEIKFFKCLYEGEITQIQSHISDTSIVLSMDNNRDLDLDSIIAEVRAQYEEIALKΞKAEAETLYQT KIQELQVTAGQHGDDLKLTKAEISELNRLIQRIRSEIGNVKKQCADLETAIADAEQRGDCALKDARAKLDELEGALHQAK EELARMLREYQELVSLKLALDMEIATYRKLLESEECRMSGEYPNSVSISVISSTNAGAGGAGFSMGFGASSSYSYKTAAA DVKTKGSCGSELKDPLAKTSGSSCATKKASR The NOVlOb amino acid sequence has 380 of 524 amino acid residues (72%) identical to, and 430 of 524 amino acid residues (82%) similar to, a Mus musculus 524 amino acid residue Type II cytokeratin (ptnr:SPTREMBL-ACC:Q9R0H5) (E = Lie^"185).

NOVlOb is expressed in at least the following tissues: skin, muscle, bone, cartilage, Colon carcinoma, lung. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Genomic Clone sources, Literature sources, and/or RACE sources. In addition, NOVlOb is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus mRNA for type II cytokeratin (GENBANK-ID: gb:GENBANK-ιD:AB033744 |acc:AB033744.1):skin, muscle, bone, cartilage, Colon carcinoma, lung.

NOVlOc

A disclosed NOV 10c nucleic acid of 1606 nucleotides (also referred to GSAC055715 B) encoding a Type II Cytokeratin-like protein is shown in Table 10E. An open reading frame was identified beginning with an ATG initiation codon at nucleotides 1-3 and ending with a TAG codon at nucleotides 1515-1517. A putative untranslated region downstream from the termination codon is underlined in Table 10E, and the start and stop codons are in bold letters.

Table 10E. NOVlOc Nucleotide Sequence (SEQ ID NO:47)

ATGAGCCGCCAATTCACCTACAAGTCGGGAGCTGCTGCCAAGGGGGGCTTCAGCGGCTGCTCCGCTGTGCTCTCAGGGGG CAGCTCATCCTCCTACCGAGCAGGGGGCAAAGGGCTCAGTGGAGGCTTCAGCAGTCGGAGCCTTTACAGCCTGGGGGGTG CCCGGAGCATCTCTTTCAATGTGGCCAGTGGCAGTGGGTGGGCAGGAGGCTATGGATTTGGCCGGGGCCGGGCCAGTGGC TTTGCTGGCAGCATGTTTGGCAGTGTGGCCTTGGGGTCCGTGTGTCCGTCGTTGTGCCCGCCCGGGGGTATCCATCAGGT CACCATCAACAAGAGCCTCCTGGCACCCCTGAACGTGGAGCTGGACCCTGAAATCCAGAAAGTGCGTGCCCAGGAGCGGG AGCAGATCAAGGTGCTGAACAACAAGTTCGCCTCCTTCATTGACAAGGTGCGGTTCCTGGAGCAGCAGAACCAGGTGCTG GAGACCAAGTGGGAGCTGCTACAGCAGCTGGACCTGAACAACTGCAAGAATAACCTGGAGCCCATCCTTGAGGGCTACAT CAGCAACCTGCGGAAGCAGCTGGAGACGCTGTCTGGGGACAGGGTGAGGCTGGACTCGGAGCTGAGGAGCGTGCGCGAAG TGGTGGAGGACTACAAGAAGAGGTATGAAGAAGAAATAAACAAGCGCACAACTGCTGAGAATGAATTTGTGGTGCTTAAG AAGGACGTGGACGCAGCTTACACGAGCAAAGTGGAGCTGCAGGCCAAGGTGGATGCCCTGGATGGAGAAATCAAGTTCTT CAAGTGTCTGTACGAGGGGGAGACTGCTCAGATCCAGTCCCACATCAGCGACACGTCCATCATCCTGTCCATGGACAACA ACCGGAACCTGGACCTGGACAGCATCATTGCTGAGGTCCGTGCCCAGTATGAGGAGATCGCCCGGAAGAGCAAGGCCGAG GCCGAGGCCCTGTACCAGACCAAGTTCCAGGAGCTGCAGCTAGCAGCCGGCCGGCATGGGGATGACCTGAAACACACCAA AAATGAGATCTCAGAGCTGACCCGTCTCATCCAAAGACTGCGCTCGGAGATTGAGAGTGTGAAGAAGCAGTGTGCCAACC TGGAGACGGCCATCGCTGACGCCGAGCAGCGGGGGGACTGTGCCCTCAAGGATGCCAGGGCCAAGCTGGATGAGCTGGAG GGCGCCCTGCAGCAGGCCAAGGAGGAGCTGGCACGGATGCTGCGCGAGTACCAAGAGCTTTTGAGCGTGAAGCTGTCCCT GGATATTGAGATCGCCACCTACCGCAAGCTGCTGGAGGGCGAGGAGTGCAGGATGTCCGGAGAATATACCAACTCCGTGA GCATTTCGGTCATCAACAGCTCCATGGCCGGGATGGCAGGCACAGGGGCTGGCTTTGGATTCAGCAATGCTGGCACCTAC GGCTACTGGCCCAGCTCTGTCAGCGGGGGCTACAGCATGCTGCCTGGGGGCTGTGTCACTGGCAGTGGGAACTGTAGCCC CCACACACACCCAGAGGGTCAGCCCCACTGGAAGTTTCCAGGGTGATCTTGGGAGTGATAACCCCAGTAATTGGAGGCCA GCAGGT The disclosed NOVlOc nucleic acid sequence, localized to chromosome 12, has 1271 of 1525 bases (83%) identical Mus musculus type II cytokeratin mRNA (gb:GENBANK-

ID:AB033744|acc:AB033744.1) (E = 5.0e^"228).

A disclosed NOVlOc polypeptide (SEQ ID NO:48) encoded by SEQ ID NO:47 is 521 amino acid residues and is presented using the one-letter amino acid code in Table 10F. Signal P, Psort and/or Hydropathy results predict that NOVlOc does not contain a signal peptide and is likely to be localized in the cytoplasm with a certainty of 0.4500.

Table 10F. Encoded NOVlOc protein sequence (SEQ ID NO:48). SRQFTYKSGAAAKGGFSGCSAVLSGGSSSSYRAGGKGLSGGFSSRSLYSLGGARSISFNVASGSG AGGYGFGRGRASG FAGSMFGSVALGSVCPSLCPPGGIHQVTINKSLLAPLNVELDPEIQKVRAQEREQIKVLNNKFASFIDKVRFLEQQNQVL ETKWELLQQLDLNNCKNNLEPILEGYISNLRKQLETLSGDRVRLDSELRSVREVVEDYKKRYEEEINKRTTAENEFVVLK KDVDAAYTSKVELQAKVDALDGEIKFFKCLYEGETAQIQΞHISDTSIILSMDNNRNLDLDSIIAEVRAQYEEIARKSKAE AEALYQTKFQELQLAAGRHGDDLKHTKNEISELTRLIQRLRSEIESVKKQCANLETAIADAEQRGDCALKDARAKLDELE GALQQAKEELARMLREYQELLSVKLSLDIEIATYRKLLEGEECRMSGEYTNSVSISVINSSMAGMAGTGAGFGFSNAGTY GYWPSSVSGGYS LPGGCVTGSGNCSPHTHPEGQPHWKFPG

The NOVlOc amino acid sequence has 414 of 506 amino acid residues (81%) identical to, and 454 of 506 amino acid residues (89%) similar to, a Mus musculus 524 amino acid residue Type II cytokeratin (ptnr:SPTREMBL-ACC:Q9R0H5) (E = ie^"212).

NOVlOc is expressed in at least the following tissues: skin, mammary gland, and lung. This information was derived by determining the tissue sources ofthe sequences that were included in the invention including but not limited to SeqCalling sources, Public EST sources, Genomic Clone sources, Literature sources, and/or RACE sources. In addition, NOVlOc is predicted to be expressed in the following tissues because ofthe expression pattern of a closely related Mus musculus mRNA for type II cytokeratin (GENBANK-ID: gb:GENBANK- ID:AB033744 |acc:AB033744.1): skin, mammary gland, and lung.

NOVlOa, NOVlOb and NOVlOc are very closely homologous as is shown in the amino acid alignment in Table 10G.

Table 10G Amino Acid Alignment of NOVlOa - NOVlOc

NOVlOa NOVlOb NOVlOb

60 70 80 90 100

NOVlOa NOVlOb NOVlOb

110 120 130 140 150 .. I .. .. I .. I .. I .. I ..| .. I

NOVlOa FGSVALGPVCPgVCPPGGIHQVTINgSLLAPLNVELDPglQKVRAQE NOVlOb FGS LGPraCPSVCPPGGlfloVTJ^NKSLLAPLNVEJajPPEIOJjjVRAOE NOVlOb MFGSVALGHVCPSESCPPGGIHOVTINKSLLAPLNVELDPEIOKVRAOE

160 170 180 190 200

I I I I I

NOVlOa REQIKALNNKFASFIDKVRFLEQQNQVLETKWELLQQLDLNNCKNNLEPI NOVlOb REQIKALNNKFASFIDKVRFLEQQNQVLETK I LLQQLDLNNC 0NLEPI NOVlOb REQIKELNNKFASFIDKVRFLEQQ_NQ_VLETKWEL_LOQLDLNNCK_NNLEPI

210 220 230 240 250

I . . I . . . . I . . I . . I . . I I -

NOVlOa LEGYISNLRKQLETLSGDRVRLDSELRNVRDVVEDYKKRYEEEINKRTAA NOVlOb I3EGYISNL|KQLE3I JLSGD@VRLDSELRN^D VEDYKKRYE2EIN|RTAA NOVlOb LEGYISNLRKQLETLSGDRVRLDSELRIVRIIVVEDYKKRYEEEINKRTØA

260 270 280 290 300 I.... I. I I . ..I .. • I

NOVlOa I 11 ID NOVlOb NOVlOb

NOVlOa NOVlOb NOVlOb

410 420 430 440 450 .. I .. .. I .. .. I .. .. I

NOVlOa JRGDJSJALKDARAKLDELEGALHQAKEELARMLREYQELgSLKLALDMEI NOVlOb )RGDCALKDARAKLDELEGALHQAKEELARMLREYQELtøSLKLALD EI NOVlOb βJjiHisHSSHBΪIi

460 470 480 490 500

NOVlOa NOVlOb NOVlOb

NOVlOa NOVlOb SR NOVlOb

Homologies to any ofthe above NOV 10 proteins will be shared by the other NOV 10 proteins insofar as they are homologous to each other as shown above. Any reference to NOV 10 is assumed to refer to both ofthe NOV 10 proteins in general, unless otherwise noted.

The disclosed NOVlOa polypeptide has homology to the amino acid sequences shown in the BLASTP data listed in Table 10H.

The homology between these and other sequences is shown graphically in the ClustalW analysis shown in Table 101. Table 101. ClustalW Analysis of NOV10

1) Novel NOVlOa (SEQ ID NO:44)

2) gj|153213021re 1XP 053295.11 (XM_053295) keratin 6 irs [Homo sapiens] (SEQ ID NO: 123)

3) gJ|15321300lref1XP 053294.11 (XM_053294) hypothetical protein XP_053294 [Homo sapiens] (SEQ ID NO: 124)

4) gil9910294lref1NP 064340.1] (NM_019956) keratin complex 2, gene 6g [Mus musculus] (SEQ ID NO: 125)

5) gi| 15321298lreflXP 053296.1 | (XM_053296) hypothetical protein XP_053296 [Homo sapiens] (SEQ ID NO: 126)

6) pi|7161776lemb|CAB76832.1| (Y19212) cytokeratin [Homo sapiens] (SEQ ID NO: 127)

60 70 80 90 100

110 120 130 140 150

210 220 230 240 250

410 420 430 440 450

Table 1 OJ lists the domain description from DOMAIN analysis results against NOVlOa. This indicates that the NOVlOa sequence has properties similar to those of other proteins known to contain these domains.

Table 10J. Domain Analysis of NOVlOa gnl | Pfam Ipfam00038, filament, Intermediate filament protein. (SEQ ID

NO:206)

Length = 312 residues, 100.0% aligned

Score = 291 bits (745), Expect = 7e-80

NOVlOa 144 QEREQIKALNNKFASFIDKVRFLEQQNQVLETKWELLQQLDLNNCKNNLEPILEGYISNL 203 l + l I++ I I++ I l + l I I I I I I I I I 1+ I I I I l + l + 1 + 1 I I

00038 NEKEQMQNLNDRLASYIDKVRFLEQQNKELEVKIEELRQ-KQAPSVSRLYSLYETEIEEL 59

NOVlOa 204 RKQLETLSGDRVRLDSELRNVRDVVEDYKKRYEEEINKRTAAENEFVLLKKDVDAAYANK 263 I + I++ I+ +I II 1+ 1 + 1+ M++I + M + IM I 111+ I l + ll + l I +

00038 60 RRQIDQLTNERARLQLEIDNLREAAEDFRKKYEDEINLRQEAENDLVGLRKDLDEATLAR 119

NOVlOa 264 VELQAKVESMDQEIKFFRCLFEAEITQIQSHISDMSVILSMDNNRNLDLDSIIDEVRTQY 323 I + I+ IMI+ +I++I + I I+ ++I+ I I +I + II I III + I + I M

00038 120 VDLENKVESLQEELEFLKKNHEEEVKELQAQIQD-TVNVEMDAARKLDLTKALREIRAQY 178

NOVlOa 324 EEIALKSKAEAEALYQTKFQELQLAAGRHGDDLKNTKNEISELTRLIQRIRSEIENVKKQ 383

MM I++ III I++I +MI II 1 + 1+ I ++ I || + || | M + I++++I I

00038 179 EEIAKKNRQEAEE YKSKLEELQTAAARNGEALRSAKEEITELRRQIQSLEIELQSLKAQ 238 NOVlOa 384 ASNLETAIADAEQRGDNALKDARAKLDELEGALHQAKEELARMLREYQELMSLKLALDME 443

++II +1+ l+l + 1+ +1 + +II I I +11+11 IIIIIII+ +IIIII+I 00038 239 NASLERQLAELEERYELELRQYQALISQLEEELQQLREEMARQLREYQELLDVKLALDIE 298

NOVlOa 444 IATYRKLLESEECR 457

II II II I II II I 00038 299 IATYRKLLEGEESR 312

Cytokeratins are constituent proteins of intermediate filaments (IFs) that form heterotypic tetrameric IF subunits containing two polypeptide chains of each ofthe two cytokeratin subfamilies, i.e. the acidic (type I) and the basic (type II). To locate the molecular domains involved in the formation of these heterotypic complexes, a binding assay was developed. The results obtained indicate that: (1) the binding between cytokeratin polypeptides ofthe complementary type is stronger and more selective than interactions of cytokeratins with other IF and non- IF proteins; (2) both the head and the tail portions ofthe proteins are not required for heterotypic complex formation; (3) the complementarity information located in the alpha-helical portions ofthe rod domain, and in short sequences immediately flanking them, is sufficient to discriminate between the two types of cytokeratins and to secure the formation of heterotypic cytokeratin complexes; (4) both coils 1 and 2 ofthe rod can contribute to this association; and (5) the formation ofthe heterotypic cytokeratin complex is not critically dependent upon ionic interactions. These results are further compatible with the concept that the heterotypic binding takes place between cytokeratin homodimer coiled-coils (Hatzfeld et al., Cytokeratin domains involved in heterotypic complex formation determined by in- vitro binding assays. J Mol Biol 197(2):237-55, 1987).

Human liver parenchymal cells have a very simple cytokeratin composition and express only one cytokeratin pair: cytokeratin 8 (a type II cytokeratin, molecular weight 52 kD) and cytokeratin 18 (a type I cytokeratin, molecular weight 45 kD). Intrahepatic bile duct cells contain in addition to cytokeratins 8 and 18 also cytokeratins 7 (a type II cytokeratin, molecular weight 54 kD) and cytokeratin 19 (a type I cytokeratin, molecular weight 40 kD) (Van Eyken et al., Immunocytochemistry of cytokeratins in primary human liver tumors. APMIS Suppl 23:77-85 1991). Three monoclonal antibodies, 1C7, 2D7 and 6B10, directed against cytokeratins of human esophagus were isolated and characterized by one- and two-dimensional gel electrophoresis and by immunohistochemical staining on sections of human epithelial tissues. In immunoblot experiments, antibodies of clones 1C7 (IgG2a) and 2D7 (IgG2b) react only with cytokeratin no. 13 ofthe acidic (type I) subfamily of cytokeratin polypeptides (Mr 54000; pi 5.1); antibodies of clone 6B10 (IgGl) detect only cytokeratin no. 4 (Mr 59000; pi 7.3) of the basic (type II) cytokeratin subfamily and allows the detection of this protein and possible degradation products at high sensitivity. These monoclonal antibodies are the first examples of antibodies specific for individual cytokeratin polypeptides characteristic of certain complex epithelia. They allow the identification of distinct minor populations of cells present in certain complex and glandular epithelia and in tumors derived therefrom which hitherto have not been distinguished. The possible reasons for the occurrence of cell type heterogeneity of cytokeratin expression in complex epithelia and in some carcinomas are discussed (van Muijen et al., Cell type heterogeneity of cytokeratin expression in complex epithelia and carcinomas as demonstrated by monoclonal antibodies specific for cytokeratins nos. 4 and 13. Exp Cell Res 162(1):97-113, 1986). Studies have analyzed the possibility that cytoskeletal proteins may be the target of forskolin in living Caco-2 cells. It was shown that forskolin initiates dramatic changes in the spatial organization ofthe cytokeratin network that correlate with an increased phosphorylation of cytokeratin molecules, whereas microtubules, micro filaments and vimentin remain mainly unaffected. Indirect immunofluorescence studies showed that the cytokeratin network is redistributed from the cell periphery to the cytoplasm. Biochemical experiments indicate that forskolin doesn't interfere with the cytokeratin profile, since the three cytokeratins normally found in intestine (CK 8, CK 18, CK 19) are similarly expressed in both control and forskolin-Caco-2 cells. Analysis of 32P-labeled cytokeratin extracted from the two cell populations demonstrates that forskolin quantitatively increases the phosphorylation of type I cytokeratin (CK 18 and CK 19), whereas the phosphorylation of type II cytokeratin (CK 8) is altered both quantitatively and qualitatively with the emergence of a new phosphorylation site (Baricault et al., The network organization and the phosphorylation of cytokeratins are concomitantly modified by forskolin in the enterocyte-like differentiated Caco-2 cell line. J Cell Sci 107 ( Pt 10):2909-18, 1994). By using the subtractive hybridization method, two complementary DNA clones differently expressed in rat normal esophageal epithelium and squamous cell carcinoma induced by administration of precursors of N-nitrososarcosine ethyl ester were isolated. A rat homologue ofthe human 50-kDa type I cytokeratin 14 was cloned for the first time and shown to be expressed preferentially in squamous cell papillomas and carcinomas, whereas it was weakly expressed or absent in normal squamous epithelial cells and in hyperplastic lesions. A rat homologue ofthe mouse 57-kDa type II cytokeratin showed strong expression in both normal and tumor tissues. These results are well consistent with the reported alteration of keratin subspecies in human esophageal cancers, therefore, encouraging us to use this experimental system as a model for human esophageal carcinogenesis (Wang et al., Identification of cytokeratin subspecies altered in rat experimental esophageal tumors by subtractive cloning. Cancer Lett 108(1): 119-27, 1996).

The protein similarity information, expression pattern, and map location for the NOV10 suggest that NOV10 may have important structural and/or physiological functions characteristic ofthe Type II Cytokeratin protein family. Therefore, the NOV10 nucleic acids and proteins ofthe invention are useful in potential therapeutic applications implicated in various diseases and disorders described below and/or other pathologies. For example, the NOV10 compositions ofthe present invention will have efficacy for treatment of patients suffering from inflammatory and infectious diseases such as AIDS; cancer, Neurologic diseases, Brain and/or autoimmune disorders like encephalomyelitis, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, endocrine diseases, muscle disorders, wound repair, bacterial, fungal, protozoal and viral infections (particularly infections caused by HIV-1 or HIV-2), pain, cancer (including but not limited to Neoplasm; adenocarcinoma; lymphoma; prostate cancer; uterus cancer), anorexia, bulimia, asthma, Parkinson's disease, acute heart failure, hypotension, hypertension, urinary retention, osteoporosis, Crohn's disease; multiple sclerosis; and Treatment of Albright Hereditary Ostoeodystrophy, angina pectoris, myocardial infarction, ulcers, asthma, allergies, benign prostatic hypertrophy, and psychotic and neurological disorders, including anxiety, schizophrenia, manic depression, delirium, dementia, severe mental retardation and dyskinesias, such as Huntington's disease. The NOV10 nucleic acid encoding Type II Cytokeratin-like protein, and the Type II Cytokeratin-like protein ofthe invention, or fragments thereof, may further be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed.

NOVX Nucleic Acids and Polypeptides One aspect ofthe invention pertains to isolated nucleic acid molecules that encode

NOVX polypeptides or biologically active portions thereof. Also included in the invention are nucleic acid fragments sufficient for use as hybridization probes to identify NOVX-encoding nucleic acids (e.g., NOVX mRNAs) and fragments for use as PCR primers for the amplification and/or mutation of NOVX nucleic acid molecules. As used herein, the term "nucleic acid molecule" is intended to include DNA molecules (e.g., cDNA or genomic DNA), RNA molecules (e.g., mRNA), analogs ofthe DNA or RNA generated using nucleotide analogs, and derivatives, fragments and homologs thereof. The nucleic acid molecule may be single-stranded or double-stranded, but preferably is comprised double- stranded DNA. An NOVX nucleic acid can encode a mature NOVX polypeptide. As used herein, a "mature" form of a polypeptide or protein disclosed in the present invention is the product of a naturally occurring polypeptide or precursor form or proprotein. The naturally occurring polypeptide, precursor or proprotein includes, by way of nonlimiting example, the full-length gene product, encoded by the corresponding gene. Alternatively, it may be defined as the polypeptide, precursor or proprotein encoded by an ORF described herein. The product "mature" form arises, again by way of nonlimiting example, as a result of one or more naturally occurring processing steps as they may take place within the cell, or host cell, in which the gene product arises. Examples of such processing steps leading to a "mature" form of a polypeptide or protein include the cleavage of the N-terminal methionine residue encoded by the initiation codon of an ORF, or the proteolytic cleavage of a signal peptide or leader sequence. Thus a mature form arising from a precursor polypeptide or protein that has residues 1 to N, where residue 1 is the N-terminal methionine, would have residues 2 through N remaining after removal ofthe N-terminal methionine. Alternatively, a mature form arising from a precursor polypeptide or protein having residues 1 to N, in which an N-terminal signal sequence from residue 1 to residue M is cleaved, would have the residues from residue M+l to residue N remaining. Further as used herein, a "mature" form of a polypeptide or protein may arise from a step of post-translational modification other than a proteolytic cleavage event. Such additional processes include, by way of non-limiting example, glycosylation, myristoylation or phosphorylation. In general, a mature polypeptide or protein may result from the operation of only one of these processes, or a combination of any of them.

The term "probes", as utilized herein, refers to nucleic acid sequences of variable length, preferably between at least about 10 nucleotides (nt), 100 nt, or as many as approximately, e.g., 6,000 nt, depending upon the specific use. Probes are used in the detection of identical, similar, or complementary nucleic acid sequences. Longer length probes are generally obtained from a natural or recombinant source, are highly specific, and much slower to hybridize than shorter-length oligomer probes. Probes may be single- or double-stranded and designed to have specificity in PCR, membrane-based hybridization technologies, or ELISA-like technologies. The term "isolated" nucleic acid molecule, as utilized herein, is one, which is separated from other nucleic acid molecules which are present in the natural source ofthe nucleic acid. Preferably, an "isolated" nucleic acid is free of sequences which naturally flank the nucleic acid (i.e., sequences located at the 5'- and 3 '-termini ofthe nucleic acid) in the genomic DNA ofthe organism from which the nucleic acid is derived. For example, in various embodiments, the isolated NOVX nucleic acid molecules can contain less than about 5 kb, 4 kb, 3 kb, 2 kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in 'genomic DNA ofthe cell/tissue from which the nucleic acid is derived (e.g., brain, heart, liver, spleen, etc.). Moreover, an "isolated" nucleic acid molecule, such as a cDNA molecule, can be substantially free of other cellular material or culture medium when produced by recombinant techniques, or of chemical precursors or other chemicals when chemically synthesized.

A nucleic acid molecule ofthe invention, e.g., a nucleic acid molecule having the nucleotide sequence SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or a complement of this aforementioned nucleotide sequence, can be isolated using standard molecular biology techniques and the sequence information provided herein. Using all or a portion ofthe nucleic acid sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 as a hybridization probe, NOVX molecules can be isolated using standard hybridization and cloning techniques (e.g. , as described in Sambrook, et αl, (eds.), MOLECULAR CLONING: A LABORATORY

MANUAL 2^nd Ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989; and Ausubel, et αl., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, NY, 1993.)

A nucleic acid ofthe invention can be amplified using cDNA, mRNA or alternatively, genomic DNA, as a template and appropriate oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to NOVX nucleotide sequences can be prepared by standard synthetic techniques, e.g., using an automated DNA synthesizer. As used herein, the term "oligonucleotide" refers to a series of linked nucleotide residues, which oligonucleotide has a sufficient number of nucleotide bases to be used in a PCR reaction. A short oligonucleotide sequence may be based on, or designed from, a genomic or cDNA sequence and is used to amplify, confirm, or reveal the presence of an identical, similar or complementary DNA or RNA in a particular cell or tissue. Oligonucleotides comprise portions of a nucleic acid sequence having about 10 nt, 50 nt, or 100 nt in length, preferably about 15 nt to 30 nt in length. In one embodiment ofthe invention, an oligonucleotide comprising a nucleic acid molecule less than 100 nt in length would further comprise at least 6 contiguous nucleotides SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or a complement thereof. Oligonucleotides may be chemically synthesized and may also be used as probes.

In another embodiment, an isolated nucleic acid molecule ofthe invention comprises a nucleic acid molecule that is a complement ofthe nucleotide sequence shown in SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or a portion of this nucleotide sequence (e.g., a fragment that can be used as a probe or primer or a fragment encoding a biologically-active portion of an NOVX polypeptide). A nucleic acid molecule that is complementary to the nucleotide sequence shown NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 or 47 is one that is sufficiently complementary to the nucleotide sequence shown NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 or 47 that it can hydrogen bond with little or no mismatches to the nucleotide sequence shown SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, thereby forming a stable duplex.

As used herein, the term "complementary" refers to Watson-Crick or Hoogsteen base pairing between nucleotides units of a nucleic acid molecule, and the term "binding" means the physical or chemical interaction between two polypeptides or compounds or associated polypeptides or compounds or combinations thereof. Binding includes ionic, non-ionic, van der Waals, hydrophobic interactions, and the like. A physical interaction can be either direct or indirect. Indirect interactions may be through or due to the effects of another polypeptide or compound. Direct binding refers to interactions that do not take place through, or due to, the effect of another polypeptide or compound, but instead are without other substantial chemical intermediates.

Fragments provided herein are defined as sequences of at least 6 (contiguous) nucleic acids or at least 4 (contiguous) amino acids, a length sufficient to allow for specific hybridization in the case of nucleic acids or for specific recognition of an epitope in the case of amino acids, respectively, and are at most some portion less than a full length sequence. Fragments may be derived from any contiguous portion of a nucleic acid or amino acid sequence of choice. Derivatives are nucleic acid sequences or amino acid sequences formed from the native compounds either directly or by modification or partial substitution. Analogs are nucleic acid sequences or amino acid sequences that have a structure similar to, but not identical to, the native compound but differs from it in respect to certain components or side chains. Analogs may be synthetic or from a different evolutionary origin and may have a similar or opposite metabolic activity compared to wild type. Homologs are nucleic acid sequences or amino acid sequences of a particular gene that are derived from different species. Derivatives and analogs may be full length or other than full length, if the derivative or analog contains a modified nucleic acid or amino acid, as described below. Derivatives or analogs ofthe nucleic acids or proteins ofthe invention include, but are not limited to, molecules comprising regions that are substantially homologous to the nucleic acids or proteins ofthe invention, in various embodiments, by at least about 70%, 80%, or 95% identity (with a preferred identity of 80-95%) over a nucleic acid or amino acid sequence of identical size or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art, or whose encoding nucleic acid is capable of hybridizing to the complement of a sequence encoding the aforementioned proteins under stringent, moderately stringent, or low stringent conditions. See e.g. Ausubel, et al., CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, New York, NY, 1993, and below.

A "homologous nucleic acid sequence" or "homologous amino acid sequence," or variations thereof, refer to sequences characterized by a homology at the nucleotide level or amino acid level as discussed above. Homologous nucleotide sequences encode those sequences coding for isoforms of NOVX polypeptides. Isoforms can be expressed in different tissues ofthe same organism as a result of, for example, alternative splicing of RNA. Alternatively, isoforms can be encoded by different genes. In the invention, homologous nucleotide sequences include nucleotide sequences encoding for an NOVX polypeptide of species other than humans, including, but not limited to: vertebrates, and thus can include, e.g., frog, mouse, rat, rabbit, dog, cat cow, horse, and other organisms. Homologous nucleotide sequences also include, but are not limited to, naturally occurring allelic variations and mutations ofthe nucleotide sequences set forth herein. A homologous nucleotide sequence does not, however, include the exact nucleotide sequence encoding human NOVX protein. Homologous nucleic acid sequences include those nucleic acid sequences that encode conservative amino acid substitutions (see below) in SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, as well as a polypeptide possessing NOVX biological activity. Various biological activities ofthe NOVX proteins are described below.

An NOVX polypeptide is encoded by the open reading frame ("ORF") of an NOVX nucleic acid. An ORF corresponds to a nucleotide sequence that could potentially be translated into a polypeptide. A stretch of nucleic acids comprising an ORF is uninterrupted by a stop codon. An ORF that represents the coding sequence for a full protein begins with an ATG "start" codon and terminates with one ofthe three "stop" codons, namely, TAA, TAG, or TGA. For the purposes of this invention, an ORF may be any part of a coding sequence, with or without a start codon, a stop codon, or both. For an ORF to be considered as a good candidate for coding for a bonafide cellular protein, a minimum size requirement is often set, e.g., a stretch of DNA that would encode a protein of 50 amino acids or more.

The nucleotide sequences determined from the cloning ofthe human NOVX genes allows for the generation of probes and primers designed for use in identifying and/or cloning NOVX homologues in other cell types, e.g. from other tissues, as well as NOVX homologues from other vertebrates. The probe/primer typically comprises substantially purified oligonucleotide. The oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, 25, 50, 100, 150, 200, 250, 300, 350 or 400 consecutive sense strand nucleotide sequence SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47; or an anti-sense strand nucleotide sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47; or of a naturally occurring mutant of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47. Probes based on the human NOVX nucleotide sequences can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In various embodiments, the probe further comprises a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a part of a diagnostic test kit for identifying cells or tissues which mis- express an NOVX protein, such as by measuring a level of an NOVX-encoding nucleic acid in a sample of cells from a subject e.g., detecting NOVX mRNA levels or determining whether a genomic NOVX gene has been mutated or deleted.

"A polypeptide having a biologically-active portion of an NOVX polypeptide" refers to polypeptides exhibiting activity similar, but not necessarily identical to, an activity of a polypeptide ofthe invention, including mature forms, as measured in a particular biological assay, with or without dose dependency. A nucleic acid fragment encoding a "biologically- active portion of NOVX" can be prepared by isolating a portion SEQ ED NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, that encodes a polypeptide having an NOVX biological activity (the biological activities ofthe NOVX proteins are described below), expressing the encoded portion of NOVX protein (e.g., by recombinant expression in vitro) and assessing the activity ofthe encoded portion of NOVX.

NOVX Nucleic Acid and Polypeptide Variants

The invention further encompasses nucleic acid molecules that differ from the nucleotide sequences shown in SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 due to degeneracy ofthe genetic code and thus encode the same NOVX proteins as that encoded by the nucleotide sequences shown in SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47. In another embodiment, an isolated nucleic acid molecule ofthe invention has a nucleotide sequence encoding a protein having an amino acid sequence shown in SEQ ID NOS:2, 4, 6, 8,

10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48.

In addition to the human NOVX nucleotide sequences shown in SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, it will be appreciated by those skilled in the art that DNA sequence polymorphisms that lead to changes in the amino acid sequences ofthe NOVX polypeptides may exist within a population (e.g., the human population). Such genetic polymorphism in the NOVX genes may exist among individuals within a population due to natural allelic variation. As used herein, the terms "gene" and "recombinant gene" refer to nucleic acid molecules comprising an open reading frame (ORF) encoding an NOVX protein, preferably a vertebrate NOVX protein. Such natural allelic variations can typically result in 1-5% variance in the nucleotide sequence ofthe NOVX genes. Any and all such nucleotide variations and resulting amino acid polymorphisms in the NOVX polypeptides, which are the result of natural allelic variation and that do not alter the functional activity ofthe NOVX polypeptides, are intended to be within the scope ofthe invention. Moreover, nucleic acid molecules encoding NOVX proteins from other species, and thus that have a nucleotide sequence that differs from the human SEQ ID NOS:l, 3, 5, 7, 9,

11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 are intended to be within the scope ofthe invention. Nucleic acid molecules corresponding to natural allelic variants and homologues ofthe NOVX cDNAs ofthe invention can be isolated based on their homology to the human NOVX nucleic acids disclosed herein using the human cDNAs, or a portion thereof, as a hybridization probe according to standard hybridization techniques under stringent hybridization conditions.

Accordingly, in another embodiment, an isolated nucleic acid molecule ofthe invention is at least 6 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47. In another embodiment, the nucleic acid is at least 10, 25, 50, 100, 250, 500, 750, 1000, 1500, or 2000 or more nucleotides in length. In yet another embodiment, an isolated nucleic acid molecule of the invention hybridizes to the coding region. As used herein, the term "hybridizes under stringent conditions" is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% homologous to each other typically remain hybridized to each other.

Homologs (t.e., nucleic acids encoding NOVX proteins derived from species other than human) or other related sequences (e.g., paralogs) can be obtained by low, moderate or high stringency hybridization with all or a portion ofthe particular human sequence as a probe using methods well known in the art for nucleic acid hybridization and cloning.

As used herein, the phrase "stringent hybridization conditions" refers to conditions under which a probe, primer or oligonucleotide will hybridize to its target sequence, but to no other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. Longer sequences hybridize specifically at higher temperatures than shorter sequences. Generally, stringent conditions are selected to be about 5 °C lower than the thermal melting point (Tm) for the specific sequence at a defined ionic strength and pH. The Tm is the temperature (under defined ionic strength, pH and nucleic acid concentration) at which 50% of the probes complementary to the target sequence hybridize to the target sequence at equilibrium. Since the target sequences are generally present at excess, at Tm, 50% ofthe probes are occupied at equilibrium. Typically, stringent conditions will be those in which the salt concentration is less than about 1.0 M sodium ion, typically about 0.01 to 1.0 M sodium ion (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes, primers or oligonucleotides (e.g., 10 nt to 50 nt) and at least about 60°C for longer probes, primers and oligonucleotides. Stringent conditions may also be achieved with the addition of destabilizing agents, such as formamide.

Stringent conditions are known to those skilled in the art and can be found in Ausubel, et al., (eds.), CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, N.Y. (1989), 6.3.1-6.3.6. Preferably, the conditions are such that sequences at least about 65%, 70%, 75%, 85%, 90%, 95%, 98%, or 99% homologous to each other typically remain hybridized to each other. A non-limiting example of stringent hybridization conditions are hybridization in a high salt buffer comprising 6X SSC, 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.02% BSA, and 500 mg/ml denatured salmon sperm DNA at 65°C, followed by one or more washes in 0.2X SSC, 0.01% BSA at 50°C. An isolated nucleic acid molecule ofthe invention that hybridizes under stringent conditions to the sequences SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, corresponds to a naturally-occurring nucleic acid molecule. As used herein, a "naturally-occurring" nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein).

In a second embodiment, a nucleic acid sequence that is hybridizable to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or fragments, analogs or derivatives thereof, under conditions of moderate stringency is provided. A non-limiting example of moderate stringency hybridization conditions are hybridization in 6X SSC, 5X Denhardt's solution, 0.5% SDS and 100 mg/ml denatured salmon sperm DNA at 55°C, followed by one or more washes in IX SSC, 0.1% SDS at 37°C. Other conditions of moderate stringency that may be used are well-known within the art. See, e.g., Ausubel, et al. (eds.),

1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990; GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY.

In a third embodiment, a nucleic acid that is hybridizable to the nucleic acid molecule comprising the nucleotide sequences SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or fragments, analogs or derivatives thereof, under conditions of low stringency, is provided. A non-limiting example of low stringency hybridization conditions are hybridization in 35% formamide, 5X SSC, 50 mM Tris-HCl (pH 7.5), 5 mM EDTA, 0.02% PVP, 0.02% Ficoll, 0.2% BSA, 100 mg/ml denatured salmon sperm DNA, 10% (wt/vol) dextran sulfate at 40°C, followed by one or more washes in 2X SSC, 25 mM Tris-HCl (pH 7.4), 5 mM EDTA, and 0.1 % SDS at 50°C. Other conditions of low stringency that may be used are well known in the art (e.g. , as employed for cross-species hybridizations). See, e.g., Ausubel, et al. (eds.), 1993, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, NY, and Kriegler, 1990, GENE TRANSFER AND EXPRESSION, A LABORATORY MANUAL, Stockton Press, NY; Shilo and Weinberg, 1981. Proc Natl Acad Sci USA 78: 6789-6792.

Conservative Mutations

In addition to naturally-occurring allelic variants of NOVX sequences that may exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation into the nucleotide sequences SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, thereby leading to changes in the amino acid sequences ofthe encoded NOVX proteins, without altering the functional ability of said NOVX proteins. For example, nucleotide substitutions leading to amino acid substitutions at "non-essential" amino acid residues can be made in the sequence SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48. A "non-essential" amino acid residue is a residue that can be altered from the wild-type sequences ofthe NOVX proteins without altering their biological activity, whereas an "essential" amino acid residue is required for such biological activity. For example, amino acid residues that are conserved among the NOVX proteins ofthe invention are predicted to be particularly non-amenable to alteration. Amino acids for which conservative substitutions can be made are well-known within the art.

Another aspect ofthe invention pertains to nucleic acid molecules encoding NOVX proteins that contain changes in amino acid residues that are not essential for activity. Such NOVX proteins differ in amino acid sequence from SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 yet retain biological activity. In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein comprises an amino acid sequence at least about 45% homologous to the amino acid sequences SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22,

24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48. Preferably, the protein encoded by the nucleic acid molecule is at least about 60% homologous to SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48; more preferably at least about 70% homologous SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48; still more preferably at least about 80% homologous to SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48; even more preferably at least about 90% homologous to SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48; and most preferably at least about 95% homologous to SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48.

An isolated nucleic acid molecule encoding an NOVX protein homologous to the protein of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48 can be created by introducing one or more nucleotide substitutions, additions or deletions into the nucleotide sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced into SEQ ID NOS: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23,

25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted, non-essential amino acid residues. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined within the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted non-essential amino acid residue in the NOVX protein is replaced with another amino acid residue from the same side chain family. Altematively, in another embodiment, mutations can be introduced randomly along all or part of an NOVX coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for NOVX biological activity to identify mutants that retain activity. Following mutagenesis SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, the encoded protein can be expressed by any recombinant technology known in the art and the activity of the protein can be determined.

The relatedness of amino acid families may also be determined based on side chain interactions. Substituted amino acids may be fully conserved "strong" residues or fully conserved "weak" residues. The "strong" group of conserved amino acid residues may be any one ofthe following groups: STA, NEQK, NHQK, NDEQ, QHRK, MILV, MILF, HY, FYW, wherein the single letter amino acid codes are grouped by those amino acids that may be substituted for each other. Likewise, the "weak" group of conserved residues may be any one ofthe following: CSA, ATV, SAG, STNK, STPA, SGND, SNDEQK, NDEQHK, NEQHRK, VLIM, HFY, wherein the letters within each group represent the single letter amino acid code. In one embodiment, a mutant NOVX protein can be assayed for (i) the ability to form proteimprotein interactions with other NOVX proteins, other cell-surface proteins, or biologically-active portions thereof, (ii) complex formation between a mutant NOVX protein and an NOVX ligand; or (ttt) the ability of a mutant NOVX protein to bind to an intracellular target protein or biologically-active portion thereof; (e.g. avidin proteins).

In yet another embodiment, a mutant NOVX protein can be assayed for the ability to regulate a specific biological function (e.g., regulation of insulin release).

Antisense Nucleic Acids

Another aspect ofthe invention pertains to isolated antisense nucleic acid molecules that are hybridizable to or complementary to the nucleic acid molecule comprising the nucleotide sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or fragments, analogs or derivatives thereof. An "antisense" nucleic acid comprises a nucleotide sequence that is complementary to a "sense" nucleic acid encoding a protein (e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence). In specific aspects, antisense nucleic acid molecules are provided that comprise a sequence complementary to at least about 10, 25, 50, 100, 250 or 500 nucleotides or an entire NOVX coding strand, or to only a portion thereof. Nucleic acid molecules encoding fragments, homologs, derivatives and analogs of an NOVX protein of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48, or antisense nucleic acids complementary to an NOVX nucleic acid sequence of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, are additionally provided.

In one embodiment, an antisense nucleic acid molecule is antisense to a "coding region" ofthe coding strand of a nucleotide sequence encoding an NOVX protein. The term "coding region" refers to the region ofthe nucleotide sequence comprising codons which are translated into amino acid residues. In another embodiment, the antisense nucleic acid molecule is antisense to a "noncoding region" ofthe coding strand of a nucleotide sequence encoding the NOVX protein. The term "noncoding region" refers to 5' and 3' sequences which flank the coding region that are not translated into amino acids (i.e., also referred to as 5' and 3' untranslated regions). Given the coding strand sequences encoding the NOVX protein disclosed herein, antisense nucleic acids ofthe invention can be designed according to the rules of Watson and Crick or Hoogsteen base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of NOVX mRNA, but more preferably is an oligonucleotide that is antisense to only a portion ofthe coding or noncoding region of NOVX mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of NOVX mRNA. An antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45 or 50 nucleotides in length. An antisense nucleic acid ofthe invention can be constructed using chemical synthesis or enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally-occurring nucleotides or variously modified nucleotides designed to increase the biological stability ofthe molecules or to increase the physical stability ofthe duplex formed between the antisense and sense nucleic acids (e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used). Examples of modified nucleotides that can be used to generate the antisense nucleic acid include: 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylaminomethyl- 2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N6-isopentenyladenine, 1-methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methylguanine, 3-methylcytosine, 5-methylcytosine, N6-adenine, 7-methylguanine, 5-methylaminomethyluracil, 5-methoxyaminomethyl-2-thiouracil, beta-D-mannosylqueosine, 5'-methoxycarboxymethyluracil, 5-methoxyuracil, 2-methylthio-N6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5-methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5-oxyacetic acid methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2-carboxypropyl) uracil, (acp3)w, and 2,6-diaminopurine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (t.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, described further in the following subsection).

The antisense nucleic acid molecules ofthe invention are typically administered to a subject or generated in situ such that they hybridize with or bind to cellular mRNA and/or genomic DNA encoding an NOVX protein to thereby inhibit expression ofthe protein (e.g., by inhibiting transcription and/or translation). The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule that binds to DNA duplexes, through specific interactions in the major groove ofthe double helix. An example of a route of administration of antisense nucleic acid molecules ofthe invention includes direct injection at a tissue site. Alternatively, antisense nucleic acid molecules can be modified to target selected cells and then administered systemically. For example, for systemic administration, antisense molecules can be modified such that they specifically bind to receptors or antigens expressed on a selected cell surface (e.g. , by linking the antisense nucleic acid molecules to peptides or antibodies that bind to cell surface receptors or antigens). The antisense nucleic acid molecules can also be delivered to cells using the vectors described herein. To achieve sufficient nucleic acid molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong pol II or pol III promoter are preferred.

In yet another embodiment, the antisense nucleic acid molecule ofthe invention is an α-anomeric nucleic acid molecule. An α-anomeric nucleic acid molecule forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual β-units, the strands run parallel to each other. See, e.g., Gaultier, et al., 1987. Nucl. Acids Res. 15: 6625-6641. The antisense nucleic acid molecule can also comprise a 2'-o-methylribonucleotide (See, e.g., Inoue, et al. 1987. Nucl. Acids Res. 15: 6131-6148) or a chimeric RNA-DNA analogue (See, e.g., Inoue, et al, 1987. FEBS Lett. 215: 327-330.

Ribozymes and PNA Moieties

Nucleic acid modifications include, by way of non- limiting example, modified bases, and nucleic acids whose sugar phosphate backbones are modified or derivatized. These modifications are carried out at least in part to enhance the chemical stability ofthe modified nucleic acid, such that they may be used, for example, as antisense binding nucleic acids in therapeutic applications in a subject.

In one embodiment, an antisense nucleic acid ofthe invention is a ribozyme. Ribozymes are catalytic RNA molecules with ribonuclease activity that are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes as described in

Haselhoff and Gerlach 1988. Nature 334: 585-591) can be used to catalytically cleave NOVX mRNA transcripts to thereby inhibit translation of NOVX mRNA. A ribozyme having specificity for an NOVX-encoding nucleic acid can be designed based upon the nucleotide sequence of an NOVX cDNA disclosed herein (i.e., SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47). For example, a derivative of a Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence ofthe active site is complementary to the nucleotide sequence to be cleaved in an NOVX-encoding mRNA. See, e.g., U.S. Patent 4,987,071 to Cech, et al. and U.S. Patent 5,116,742 to Cech, et al. NOVX mRNA can also be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules. See, e.g., Bartel et al, (1993) Science 261 :1411-1418. Alternatively, NOVX gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region ofthe NOVX nucleic acid (e.g., the NOVX promoter and/or enhancers) to form triple helical structures that prevent transcription ofthe NOVX gene in target cells. See, e.g., Helene, 1991. Anticancer Drug Des. 6: 569-84; Helene, et al. 1992. Ann. N.Y. Acad. Sci. 660: 27-36; Maher, 1992. Bioassays 14: 807-15.

In various embodiments, the NOVX nucleic acids can be modified at the base moiety, sugar moiety or phosphate backbone to improve, e.g., the stability, hybridization, or solubility ofthe molecule. For example, the deoxyribose phosphate backbone ofthe nucleic acids can be modified to generate peptide nucleic acids. See, e.g., Hyrup, et al., 1996. Bioorg Med Chem 4: 5-23. As used herein, the terms "peptide nucleic acids" or "PNAs" refer to nucleic acid mimics (e.g., DNA mimics) in which the deoxyribose phosphate backbone is replaced by a pseudopeptide backbone and only the four natural nucleobases are retained. The neutral backbone of PNAs has been shown to allow for specific hybridization to DNA and RNA under conditions of low ionic strength. The synthesis of PNA oligomers can be performed using standard solid phase peptide synthesis protocols as described in Hyrup, et ai, 1996. supra; Perry-O'Keefe, et al., 1996. Proc. Natl. Acad. Sci. USA 93: 14670-14675.

PNAs of NOVX can be used in therapeutic and diagnostic applications. For example, PNAs can be used as antisense or antigene agents for sequence-specific modulation of gene expression by, e.g., inducing transcription or translation arrest or inhibiting replication. PNAs of NOVX can also be used, for example, in the analysis of single base pair mutations in a gene (e.g. , PNA directed PCR clamping; as artificial restriction enzymes when used in combination with other enzymes, e.g., Si nucleases (See, Hyrup, et al., 1996.supra); or as probes or primers for DNA sequence and hybridization (See, Hyrup, et al., 1996, supra; Perry-O'Keefe, et al., 1996. supra).

In another embodiment, PNAs of NOVX can be modified, e.g., to enhance their stability or cellular uptake, by attaching lipophilic or other helper groups to PNA, by the formation of PNA-DNA chimeras, or by the use of liposomes or other techniques of drug delivery known in the art. For example, PNA-DNA chimeras of NOVX can be generated that may combine the advantageous properties of PNA and DNA. Such chimeras allow DNA recognition enzymes (e.g., RNase H and DNA polymerases) to interact with the DNA portion while the PNA portion would provide high binding affinity and specificity. PNA-DNA chimeras can be linked using linkers of appropriate lengths selected in terms of base stacking, number of bonds between the nucleobases, and orientation (see, Hyrup, et al., 1996. supra). The synthesis of PNA-DNA chimeras can be performed as described in Hyrup, et al., 1996. supra and Finn, et ai, 1996. Nucl Acids Res 24: 3357-3363. For example, a DNA chain can be synthesized on a solid support using standard phosphoramidite coupling chemistry, and modified nucleoside analogs, e.g., 5'-(4-methoxytrityl)amino-5'-deoxy-thymidine phosphoramidite, can be used between the PNA and the 5' end of DNA. See, e.g., Mag, et al., 1989. Nucl Acid Res 17: 5973-5988. PNA monomers are then coupled in a stepwise manner to produce a chimeric molecule with a 5' PNA segment and a 3' DNA segment. See, e.g., Finn, et al., 1996. supra. Alternatively, chimeric molecules can be synthesized with a 5' DNA segment and a 3' PNA segment. See, e.g., Petersen, et al., 1975. Bioorg. Med. Chem. Lett. 5: 1119-11124. In other embodiments, the oligonucleotide may include other appended groups such as peptides (e.g. , for targeting host cell receptors in vivo), or agents facilitating transport across the cell membrane (see, e.g., Letsinger, et al., 1989. Proc. Natl. Acad. Sci. U.S.A. 86: 6553-6556; Lemaitre, et al., 1987. Proc. Natl. Acad. Sci. 84: 648-652; PCT Publication No. WO88/09810) or the blood-brain barrier (see, e.g., PCT Publication No. WO 89/10134). In addition, oligonucleotides can be modified with hybridization triggered cleavage agents (see, e.g., Krol, et al., 1988. BioTechniques 6:958-976) or intercalating agents (see, e.g., Zon, 1988. Pharm. Res. 5: 539-549). To this end, the oligonucleotide may be conjugated to another molecule, e.g. , a peptide, a hybridization triggered cross-linking agent, a transport agent, a hybridization-triggered cleavage agent, and the like.

NOVX Polypeptides

A polypeptide according to the invention includes a polypeptide including the amino acid sequence of NOVX polypeptides whose sequences are provided in SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48. The invention also includes a mutant or variant protein any of whose residues may be changed from the corresponding residues shown in SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48 while still encoding a protein that maintains its NOVX activities and physiological functions, or a functional fragment thereof. In general, an NOVX variant that preserves NOVX-like function includes any variant in which residues at a particular position in the sequence have been substituted by other amino acids, and further include the possibility of inserting an additional residue or residues between two residues ofthe parent protein as well as the possibility of deleting one or more residues from the parent sequence. Any amino acid substitution, insertion, or deletion is encompassed by the invention. In favorable circumstances, the substitution is a conservative substitution as defined above.

One aspect ofthe invention pertains to isolated NOVX proteins, and biologically- active portions thereof, or derivatives, fragments, analogs or homologs thereof. Also provided are polypeptide fragments suitable for use as immunogens to raise anti-NOVX antibodies. In one embodiment, native NOVX proteins can be isolated from cells or tissue sources by an appropriate purification scheme using standard protein purification techniques. In another embodiment, NOVX proteins are produced by recombinant DNA techniques. Alternative to recombinant expression, an NOVX protein or polypeptide can be synthesized chemically using standard peptide synthesis techniques. An "isolated" or "purified" polypeptide or protein or biologically-active portion thereof is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the NOVX protein is derived, or substantially free from chemical precursors or other chemicals when chemically synthesized. The language "substantially free of cellular material" includes preparations of NOVX proteins in which the protein is separated from cellular components ofthe cells from which it is isolated or recombinantly-produced. In one embodiment, the language "substantially free of cellular material" includes preparations of NOVX proteins having less than about 30% (by dry weight) of non-NOVX proteins (also referred to herein as a "contaminating protein"), more preferably less than about 20% of non-NOVX proteins, still more preferably less than about 10% of non-NOVX proteins, and most preferably less than about 5% of non-NOVX proteins. When the NOVX protein or biologically-active portion thereof is recombinantly-produced, it is also preferably substantially free of culture medium, t.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% ofthe volume of the NOVX protein preparation.

The language "substantially free of chemical precursors or other chemicals" includes preparations of NOVX proteins in which the protein is separated from chemical precursors or other chemicals that are involved in the synthesis ofthe protein. In one embodiment, the language "substantially free of chemical precursors or other chemicals" includes preparations of NOVX proteins having less than about 30% (by dry weight) of chemical precursors or non-NOVX chemicals, more preferably less than about 20% chemical precursors or non-NOVX chemicals, still more preferably less than about 10% chemical precursors or non-NOVX chemicals, and most preferably less than about 5% chemical precursors or non-NOVX chemicals. Biologically-active portions of NOVX proteins include peptides comprising amino acid sequences sufficiently homologous to or derived from the amino acid sequences ofthe NOVX proteins (e.g., the amino acid sequence shown in SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48) that include fewer amino acids than the full-length NOVX proteins, and exhibit at least one activity of an NOVX protein. Typically, biologically-active portions comprise a domain or motif with at least one activity ofthe NOVX protein. A biologically-active portion of an NOVX protein can be a polypeptide which is, for example, 10, 25, 50, 100 or more amino acid residues in length. Moreover, other biologically-active portions, in which other regions ofthe protein are deleted, can be prepared by recombinant techniques and evaluated for one or more ofthe functional activities of a native NOVX protein.

In an embodiment, the NOVX protein has an amino acid sequence shown SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48. In other embodiments, the NOVX protein is substantially homologous to SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48, and retains the functional activity ofthe protein of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48, yet differs in amino acid sequence due to natural allelic variation or mutagenesis, as described in detail, below. Accordingly, in another embodiment, the NOVX protein is a protein that comprises an amino acid sequence at least about 45% homologous to the amino acid sequence SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48, and retains the functional activity ofthe NOVX proteins of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48.

Determining Homology Between Two or More Sequences

To determine the percent homology of two amino acid sequences or of two nucleic acids, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first amino acid or nucleic acid sequence for optimal alignment with a second amino or nucleic acid sequence). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence, then the molecules are homologous at that position (t.e., as used herein amino acid or nucleic acid "homology" is equivalent to amino acid or nucleic acid "identity").

The nucleic acid sequence homology may be determined as the degree of identity between two sequences. The homology may be determined using computer programs known in the art, such as GAP software provided in the GCG program package. See, Needleman and Wunsch, 1970. J Mol Biol 48: 443-453. Using GCG GAP software with the following settings for nucleic acid sequence comparison: GAP creation penalty of 5.0 and GAP extension penalty of 0.3, the coding region ofthe analogous nucleic acid sequences referred to above exhibits a degree of identity preferably of at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%, with the CDS (encoding) part ofthe DNA sequence shown in SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47. The term "sequence identity" refers to the degree to which two polynucleotide or polypeptide sequences are identical on a residue-by-residue basis over a particular region of comparison. The term "percentage of sequence identity" is calculated by comparing two optimally aligned sequences over that region of comparison, determining the number of positions at which the identical nucleic acid base (e.g., A, T, C, G, U, or I, in the case of nucleic acids) occurs in both sequences to yield the number of matched positions, dividing the number of matched positions by the total number of positions in the region of comparison (t.e., the window size), and multiplying the result by 100 to yield the percentage of sequence identity. The term "substantial identity" as used herein denotes a characteristic of a polynucleotide sequence, wherein the polynucleotide comprises a sequence that has at least 80 percent sequence identity, preferably at least 85 percent identity and often 90 to 95 percent sequence identity, more usually at least 99 percent sequence identity as compared to a reference sequence over a comparison region.

Chimeric and Fusion Proteins The invention also provides NOVX chimeric or fusion proteins. As used herein, an

NOVX "chimeric protein" or "fusion protein" comprises an NOVX polypeptide operatively- linked to a non-NOVX polypeptide. An "NOVX polypeptide" refers to a polypeptide having an amino acid sequence corresponding to an NOVX protein SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and 48, whereas a "non-NOVX polypeptide" refers to a polypeptide having an amino acid sequence corresponding to a protein that is not substantially homologous to the NOVX protein, e.g., a protein that is different from the NOVX protein and that is derived from the same or a different organism. Within an NOVX fusion protein the NOVX polypeptide can correspond to all or a portion of an NOVX protein. In one embodiment, an NOVX fusion protein comprises at least one biologically- active portion of an NOVX protein. In another embodiment, an NOVX fusion protein comprises at least two biologically-active portions of an NOVX protein. In yet another embodiment, an NOVX fusion protein comprises at least three biologically-active portions of an NOVX protein. Within the fusion protein, the term "operatively-linked" is intended to indicate that the NOVX polypeptide and the non-NOVX polypeptide are fused in- frame with one another. The non-NOVX polypeptide can be fused to the N-terminus or C-terminus ofthe NOVX polypeptide.

In one embodiment, the fusion protein is a GST-NOVX fusion protein in which the NOVX sequences are fused to the C-terminus ofthe GST (glutathione S -transferase) sequences. Such fusion proteins can facilitate the purification of recombinant NOVX polypeptides.

In another embodiment, the fusion protein is an NOVX protein containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression and/or secretion of NOVX can be increased through use of a heterologous signal sequence.

In yet another embodiment, the fusion protein is an NOVX-immunoglobulin fusion protein in which the NOVX sequences are fused to sequences derived from a member ofthe immunoglobulin protein family. The NOVX-immunoglobulin fusion proteins ofthe invention can be incorporated into pharmaceutical compositions and administered to a subject to inhibit an interaction between an NOVX ligand and an NOVX protein on the surface of a cell, to thereby suppress NOVX-mediated signal transduction in vivo. The NOVX-immunoglobulin fusion proteins can be used to affect the bioavailability of an NOVX cognate ligand. Inhibition ofthe NOVX ligand/NOVX interaction may be useful therapeutically for both the treatment of proliferative and differentiative disorders, as well as modulating (e.g. promoting or inhibiting) cell survival. Moreover, the NOVX-immunoglobulin fusion proteins ofthe invention can be used as immunogens to produce anti-NOVX antibodies in a subject, to purify NOVX ligands, and in screening assays to identify molecules that inhibit the interaction of NOVX with an NOVX ligand. An NOVX chimeric or fusion protein ofthe invention can be produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, e.g., by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments that can subsequently be annealed and reamplified to generate a chimeric gene sequence (see, e.g., Ausubel, et al. (eds.) CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, John Wiley & Sons, 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). An NOVX-encoding nucleic acid can be cloned into such an expression vector such that the fusion moiety is linked in-frame to the NOVX protein. NOVX Agonists and Antagonists

The invention also pertains to variants ofthe NOVX proteins that function as either NOVX agonists (i.e., mimetics) or as NOVX antagonists. Variants ofthe NOVX protein can be generated by mutagenesis (e.g., discrete point mutation or truncation ofthe NOVX protein). An agonist ofthe NOVX protein can retain substantially the same, or a subset of, the biological activities ofthe naturally occurring form ofthe NOVX protein. An antagonist of the NOVX protein can inhibit one or more ofthe activities ofthe naturally occurring form of the NOVX protein by, for example, competitively binding to a downstream or upstream member of a cellular signaling cascade which includes the NOVX protein. Thus, specific biological effects can be elicited by treatment with a variant of limited function. In one embodiment, treatment of a subject with a variant having a subset ofthe biological activities ofthe naturally occurring form ofthe protein has fewer side effects in a subject relative to treatment with the naturally occurring form ofthe NOVX proteins.

Variants ofthe NOVX proteins that function as either NOVX agonists (i.e., mimetics) or as NOVX antagonists can be identified by screening combinatorial libraries of mutants (e.g., truncation mutants) ofthe NOVX proteins for NOVX protein agonist or antagonist activity. In one embodiment, a variegated library of NOVX variants is generated by combinatorial mutagenesis at the nucleic acid level and is encoded by a variegated gene library. A variegated library of NOVX variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential NOVX sequences is expressible as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g. , for phage display) containing the set of NOVX sequences therein. There are a variety of methods which can be used to produce libraries of potential NOVX variants from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene then ligated into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all ofthe sequences encoding the desired set of potential NOVX sequences. Methods for synthesizing degenerate oligonucleotides are well-known within the art. See, e.g., Narang, 1983. Tetrahedron 39: 3; Itakura, et al., 1984. Annu. Rev. Biochem. 53: 323; Itakura, et al., 1984. Science 198: 1056; Ike, et al., 1983. Nucl. Acids Res. 11: 477.

Polypeptide Libraries

In addition, libraries of fragments ofthe NOVX protein coding sequences can be used to generate a variegated population of NOVX fragments for screening and subsequent selection of variants of an NOVX protein. In one embodiment, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of an NOVX coding sequence with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, renaturing the DNA to form double-stranded DNA that can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with Si nuclease, and ligating the resulting fragment library into an expression vector. By this method, expression libraries can be derived which encodes N-terminal and internal fragments of various sizes ofthe NOVX proteins. Various techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. Such techniques are adaptable for rapid screening ofthe gene libraries generated by the combinatorial mutagenesis of NOVX proteins. The most widely used techniques, which are amenable to high throughput analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation ofthe vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a new technique that enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify NOVX variants. See, e.g., Arkin and Yourvan, 1992. Proc. Natl. Acad. Sci. USA 89: 7811-7815; Delgrave, et al., 1993. Protein Engineering 6:327-331.

Anti-NOVX Antibodies

Also included in the invention are antibodies to NOVX proteins, or fragments of NOVX proteins. The term "antibody" as used herein refers to immunoglobulin molecules and immunologically active portions of immunoglobulin (Ig) molecules, i.e., molecules that contain an antigen binding site that specifically binds (immunoreacts with) an antigen. Such antibodies include, but are not limited to, polyclonal, monoclonal, chimeric, single chain, F_ab, Fab' and F(_ab-)2 fragments, and an F_ab expression library. In general, an antibody molecule obtained from humans relates to any ofthe classes IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature ofthe heavy chain present in the molecule. Certain classes have subclasses as well, such as IgGi, IgG₂, and others. Furthermore, in humans, the light chain may be a kappa chain or a lambda chain. Reference herein to antibodies includes a reference to all such classes, subclasses and types of human antibody species. An isolated NOVX-related protein ofthe invention may be intended to serve as an antigen, or a portion or fragment thereof, and additionally can be used as an immunogen to generate antibodies that immunospecifically bind the antigen, using standard techniques for polyclonal and monoclonal antibody preparation. The full-length protein can be used or, alternatively, the invention provides antigenic peptide fragments ofthe antigen for use as immunogens. An antigenic peptide fragment comprises at least 6 amino acid residues ofthe amino acid sequence ofthe full length protein and encompasses an epitope thereof such that an antibody raised against the peptide forms a specific immune complex with the full length protein or with any fragment that contains the epitope. Preferably, the antigenic peptide comprises at least 10 amino acid residues, or at least 15 amino acid residues, or at least 20 amino acid residues, or at least 30 amino acid residues. Preferred epitopes encompassed by the antigenic peptide are regions ofthe protein that are located on its surface; commonly these are hydrophilic regions.

In certain embodiments ofthe invention, at least one epitope encompassed by the antigenic peptide is a region of NOVX-related protein that is located on the surface ofthe protein, e.g., a hydrophilic region. A hydrophobicity analysis ofthe human NOVX-related protein sequence will indicate which regions of a NOVX-related protein are particularly hydrophilic and, therefore, are likely to encode surface residues useful for targeting antibody production. As a means for targeting antibody production, hydropathy plots showing regions of hydrophilicity and hydrophobicity may be generated by any method well known in the art, including, for example, the Kyte Doolittle or the Hopp Woods methods, either with or without Fourier transformation. See, e.g., Hopp and Woods, 1981, Proc. Nat. Acad. Sci. USA 78: 3824-3828; Kyte and Doolittle 1982, J. Mol. Biol. 157: 105-142, each of which is incorporated herein by reference in its entirety. Antibodies that are specific for one or more domains within an antigenic protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

A protein ofthe invention, or a derivative, fragment, analog, homolog or ortho log thereof, may be utilized as an immunogen in the generation of antibodies that immunospecifically bind these protein components. Various procedures known within the art may be used for the production of polyclonal or monoclonal antibodies directed against a protein ofthe invention, or against derivatives, fragments, analogs homologs or orthologs thereof (see, for example, Antibodies: A Laboratory Manual, Harlow and Lane, 1988, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, incorporated herein by reference). Some of these antibodies are discussed below. Polyclonal Antibodies

For the production of polyclonal antibodies, various suitable host animals (e.g., rabbit, goat, mouse or other mammal) may be immunized by one or more injections with the native protein, a synthetic variant thereof, or a derivative ofthe foregoing. An appropriate immunogenic preparation can contain, for example, the naturally occurring immunogenic protein, a chemically synthesized polypeptide representing the immunogenic protein, or a recombinantly expressed immunogenic protein. Furthermore, the protein may be conjugated to a second protein known to be immunogenic in the mammal being immunized. Examples of such immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor. The preparation can further include an adjuvant. Various adjuvants used to increase the immunological response include, but are not limited to, Freund's (complete and incomplete), mineral gels (e.g., aluminum hydroxide), surface active substances (e.g., lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, dinitrophenol, etc.), adjuvants usable in humans such as Bacille Calmette-Guerin and Corynebacterium parvum, or similar immunostimulatory agents. Additional examples of adjuvants which can be employed include MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).

The polyclonal antibody molecules directed against the immunogenic protein can be isolated from the mammal (e.g., from the blood) and further purified by well known techniques, such as affinity chromatography using protein A or protein G, which provide primarily the IgG fraction of immune serum. Subsequently, or alternatively, the specific antigen which is the target ofthe immunoglobulin sought, or an epitope thereof, may be immobilized on a column to purify the immune specific antibody by immunoaffmity chromatography. Purification of immunoglobulins is discussed, for example, by D. Wilkinson (The Scientist, published by The Scientist, Inc., Philadelphia PA, Vol. 14, No. 8 (April 17, 2000), pp. 25-28).

Monoclonal Antibodies

The term "monoclonal antibody" (MAb) or "monoclonal antibody composition", as used herein, refers to a population of antibody molecules that contain only one molecular species of antibody molecule consisting of a unique light chain gene product and a unique heavy chain gene product. In particular, the complementarity determining regions (CDRs) of the monoclonal antibody are identical in all the molecules ofthe population. MAbs thus contain an antigen binding site capable of immunoreacting with a particular epitope ofthe antigen characterized by a unique binding affinity for it. Monoclonal antibodies can be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature, 256:495 (1975). In a hybridoma method, a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent. Alternatively, the lymphocytes can be immunized in vitro.

The immunizing agent will typically include the protein antigen, a fragment thereof or a fusion protein thereof. Generally, either peripheral blood lymphocytes are used if cells of human origin are desired, or spleen cells or lymph node cells are used if non-human mammalian sources are desired. The lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell

(Goding, MONOCLONAL ANTIBODIES: PRINCIPLES AND PRACTICE, Academic Press, (1986) pp. 59-103). Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin. Usually, rat or mouse myeloma cell lines are employed. The hybridoma cells can be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells. For example, if the parental cells lack the enzyme hypoxanthine guanine phosphoribosyl transferase (HGPRT or HPRT), the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine ("HAT medium"), which substances prevent the growth of HGPRT-deficient cells. Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, California and the American Type Culture Collection, Manassas, Virginia. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies (Kozbor, J. Immunol, 133:3001 (1984); Brodeur et al., MONOCLONAL ANTIBODY PRODUCTION TECHNIQUES AND APPLICATIONS, Marcel Dekker, Inc., New York, (1987) pp. 51-63).

The culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against the antigen. Preferably, the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA). Such techniques and assays are known in the art. The binding affinity ofthe monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem., 107:220 (1980). Preferably, antibodies having a high degree of specificity and a high binding affinity for the target antigen are isolated.

After the desired hybridoma cells are identified, the clones can be subcloned by limiting dilution procedures and grown by standard methods. Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium. Alternatively, the hybridoma cells can be grown in vivo as ascites in a mammal.

The monoclonal antibodies secreted by the subclones can be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.

The monoclonal antibodies can also be made by recombinant DNA methods, such as those described in U.S. Patent No. 4,816,567. DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies). The hybridoma cells ofthe invention serve as a preferred source of such DNA. Once isolated, the DNA can be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells. The DNA also can be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place ofthe homologous murine sequences (U.S. Patent No. 4,816,567; Morrison, Nature 368, 812-13 (1994)) or by covalently joining to the immunoglobulin coding sequence all or part ofthe coding sequence for a non-immunoglobulin polypeptide. Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody ofthe invention, or can be substituted for the variable domains of one antigen-combining site of an antibody ofthe invention to create a chimeric bivalent antibody.

Humanized Antibodies

The antibodies directed against the protein antigens ofthe invention can further comprise humanized antibodies or human antibodies. These antibodies are suitable for administration to humans without engendering an immune response by the human against the administered immunoglobulin. Humanized forms of antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab')₂ or other antigen- binding subsequences of antibodies) that are principally comprised ofthe sequence of a human immunoglobulin, and contain minimal sequence derived from a non-human immunoglobulin. Humanization can be performed following the method of Winter and co-workers (Jones et al., Nature, 321 :522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science, 239:1534-1536 (1988)), by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody. (See also U.S. Patent No. 5,225,539.) In some instances, Fv framework residues ofthe human immunoglobulin are replaced by corresponding non-human residues. Humanized antibodies can also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences. In general, the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all ofthe CDR regions correspond to those of a non-human immunoglobulin and all or substantially all ofthe framework regions are those of a human immunoglobulin consensus sequence. The humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin (Jones et al., 1986; Riechmann et al., 1988; and Presta, Curr. Op. Struct. Biol, 2:593-596 (1992)).

Human Antibodies

Fully human antibodies relate to antibody molecules in which essentially the entire sequences of both the light chain and the heavy chain, including the CDRs, arise from human genes. Such antibodies are termed "human antibodies", or "fully human antibodies" herein. Human monoclonal antibodies can be prepared by the trioma technique; the human B-cell hybridoma technique (see Kozbor, et al., 1983 Immunol Today 4: 72) and the EBV hybridoma technique to produce human monoclonal antibodies (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96). Human monoclonal antibodies may be utilized in the practice ofthe present invention and may be produced by using human hybridomas (see Cote, et al., 1983. Proc Natl Acad Sci USA 80: 2026-2030) or by transforming human B-cells with Epstein Barr Virus in vitro (see Cole, et al., 1985 In: MONOCLONAL ANTIBODIES AND CANCER THERAPY, Alan R. Liss, Inc., pp. 77-96).

In addition, human antibodies can also be produced using additional techniques, including phage display libraries (Hoogenboom and Winter, J. Mol. Biol., 227:381 (1991); Marks et al., J. Mol. Biol, 222:581 (1991)). Similarly, human antibodies can be made by introducing human immunoglobulin loci into transgenic animals, e.g., mice in which the endogenous immunoglobulin genes have been partially or completely inactivated. Upon challenge, human antibody production is observed, which closely resembles that seen in humans in all respects, including gene rearrangement, assembly, and antibody repertoire. This approach is described, for example, in U.S. Patent Nos. 5,545,807; 5,545,806; 5,569,825; 5,625,126; 5,633,425; 5,661,016, and in Marks et al. (Bio/Technology 10, 779-783 (1992)); Lonberg et al. (Nature 368 856-859 (1994)); Morrison ( Nature 368, 812-13 (1994)); Fishwild et al,( Nature Biotechnology 14, 845-51 (1996)); Neuberger (Nature Biotechnology 14, 826 (1996)); and Lonberg and Huszar (Intern. Rev. Immunol. 13 65-93 (1995)).

Human antibodies may additionally be produced using transgenic nonhuman animals which are modified so as to produce fully human antibodies rather than the animal's endogenous antibodies in response to challenge by an antigen. (See PCT publication WO94/02602). The endogenous genes encoding the heavy and light immunoglobulin chains in the nonhuman host have been incapacitated, and active loci encoding human heavy and light chain immunoglobulins are inserted into the host's genome. The human genes are incorporated, for example, using yeast artificial chromosomes containing the requisite human DNA segments. An animal which provides all the desired modifications is then obtained as progeny by crossbreeding intermediate transgenic animals containing fewer than the full complement ofthe modifications. The preferred embodiment of such a nonhuman animal is a mouse, and is termed the Xenomouse™ as disclosed in PCT publications WO 96/33735 and WO 96/34096. This animal produces B cells which secrete fully human immunoglobulins. The antibodies can be obtained directly from the animal after immunization with an immunogen of interest, as, for example, a preparation of a polyclonal antibody, or alternatively from immortalized B cells derived from the animal, such as hybridomas producing monoclonal antibodies. Additionally, the genes encoding the immunoglobulins with human variable regions can be recovered and expressed to obtain the antibodies directly, or can be further modified to obtain analogs of antibodies such as, for example, single chain Fv molecules. An example of a method of producing a nonhuman host, exemplified as a mouse, lacking expression of an endogenous immunoglobulin heavy chain is disclosed in U.S. Patent No. 5,939,598. It can be obtained by a method including deleting the J segment genes from at least one endogenous heavy chain locus in an embryonic stem cell to prevent rearrangement of the locus and to prevent formation of a transcript of a rearranged immunoglobulin heavy chain locus, the deletion being effected by a targeting vector containing a gene encoding a selectable marker; and producing from the embryonic stem cell a transgenic mouse whose somatic and germ cells contain the gene encoding the selectable marker.

A method for producing an antibody of interest, such as a human antibody, is disclosed in U.S. Patent No. 5,916,771. It includes introducing an expression vector that contains a nucleotide sequence encoding a heavy chain into one mammalian host cell in culture, introducing an expression vector containing a nucleotide sequence encoding a light chain into another mammalian host cell, and fusing the two cells to form a hybrid cell. The hybrid cell expresses an antibody containing the heavy chain and the light chain. In a further improvement on this procedure, a method for identifying a clinically relevant epitope on an immunogen, and a correlative method for selecting an antibody that binds immunospecifically to the relevant epitope with high affinity, are disclosed in PCT publication WO 99/53049.

F_ab Fragments and Single Chain Antibodies According to the invention, techniques can be adapted for the production of single-chain antibodies specific to an antigenic protein ofthe invention (see e.g., U.S. Patent No. 4,946,778). In addition, methods can be adapted for the construction of F_ab expression libraries (see e.g., Huse, et al., 1989 Science 246: 1275-1281) to allow rapid and effective identification of monoclonal F_ab fragments with the desired specificity for a protein or derivatives, fragments, analogs or homologs thereof. Antibody fragments that contain the idiotypes to a protein antigen may be produced by techniques known in the art including, but not limited to: (i) an F(_{ab )2} fragment produced by pepsin digestion of an antibody molecule; (ii) an F_ab fragment generated by reducing the disulfide bridges of an F_(ab')2 fragment; (iii) an F_ab fragment generated by the treatment ofthe antibody molecule with papain and a reducing agent and (iv) F_v fragments.

Bispecific Antibodies

Bispecific antibodies are monoclonal, preferably human or humanized, antibodies that have binding specificities for at least two different antigens. In the present case, one ofthe binding specificities is for an antigenic protein ofthe invention. The second binding target is any other antigen, and advantageously is a cell-surface protein or receptor or receptor subunit. Methods for making bispecific antibodies are known in the art. Traditionally, the recombinant production of bispecific antibodies is based on the co-expression of two immunoglobulin heavy-chain/light-chain pairs, where the two heavy chains have different specificities (Milstein and Cuello, Nature, 305:537-539 (1983)). Because ofthe random assortment of immunoglobulin heavy and light chains, these hybridomas (quadromas) produce a potential mixture often different antibody molecules, of which only one has the correct bispecific structure. The purification ofthe correct molecule is usually accomplished by affinity chromatography steps. Similar procedures are disclosed in WO 93/08829, published 13 May 1993, and in Traunecker et al, 1991 EMBOJ., 10:3655-3659.

Antibody variable domains with the desired binding specificities (antibody-antigen combining sites) can be fused to immunoglobulin constant domain sequences. The fusion preferably is with an immunoglobulin heavy-chain constant domain, comprising at least part ofthe hinge, CH2, and CH3 regions. It is preferred to have the first heavy-chain constant region (CHI) containing the site necessary for light-chain binding present in at least one ofthe fusions. DNAs encoding the immunoglobulin heavy-chain fusions and, if desired, the immunoglobulin light chain, are inserted into separate expression vectors, and are co- transfected into a suitable host organism. For further details of generating bispecific antibodies see, for example, Suresh et al., Methods in Enzymology, 121:210 (1986).

According to another approach described in WO 96/27011, the interface between a pair of antibody molecules can be engineered to maximize the percentage of heterodimers which are recovered from recombinant cell culture. The preferred interface comprises at least a part ofthe CH3 region of an antibody constant domain. In this method, one or more small amino acid side chains from the interface ofthe first antibody molecule are replaced with larger side chains (e.g. tyrosine or tryptophan). Compensatory "cavities" of identical or similar size to the large side chain(s) are created on the interface ofthe second antibody molecule by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). This provides a mechanism for increasing the yield ofthe heterodimer over other unwanted end-products such as homodimers.

Bispecific antibodies can be prepared as full length antibodies or antibody fragments (e.g. F(ab')₂ bispecific antibodies). Techniques for generating bispecific antibodies from antibody fragments have been described in the literature. For example, bispecific antibodies can be prepared using chemical linkage. Brennan et al., Science 229:81 (1985) describe a procedure wherein intact antibodies are proteolytic ally cleaved to generate F(ab')₂ fragments. These fragments are reduced in the presence ofthe dithiol complexing agent sodium arsenite to stabilize vicinal dithiols and prevent intermolecular disulfide formation. The Fab' fragments generated are then converted to thionitrobenzoate (TNB) derivatives. One ofthe Fab' -TNB derivatives is then reconverted to the Fab '-thiol by reduction with mercaptoethylamine and is mixed with an equimolar amount ofthe other Fab'-TNB derivative to form the bispecific antibody. The bispecific antibodies produced can be used as agents for the selective immobilization of enzymes. Additionally, Fab' fragments can be directly recovered from E. coli and chemically coupled to form bispecific antibodies. Shalaby et al., J. Exp. Med. 175:217-225 (1992) describe the production of a fully humanized bispecific antibody F(ab')₂ molecule. Each Fab' fragment was separately secreted from E. coli and subjected to directed chemical coupling in vitro to form the bispecific antibody. The bispecific antibody thus formed was able to bind to cells overexpressing the ErbB2 receptor and normal human T cells, as well as trigger the lytic activity of human cytotoxic lymphocytes against human breast tumor targets.

Various techniques for making and isolating bispecific antibody fragments directly from recombinant cell culture have also been described. For example, bispecific antibodies have been produced using leucine zippers. Kostelny et al., J. Immunol. 148(5):1547-1553 (1992). The leucine zipper peptides from the Fos and Jun proteins were linked to the Fab' portions of two different antibodies by gene fusion. The antibody homodimers were reduced at the hinge region to form monomers and then re-oxidized to form the antibody heterodimers. This method can also be utilized for the production of antibody homodimers. The "diabody" technology described by Hollinger et al., Proc. Natl. Acad. Sci. USA 90:6444-6448 (1993) has provided an alternative mechanism for making bispecific antibody fragments. The fragments comprise a heavy-chain variable domain (V_H) connected to a light-chain variable domain (V_L) by a linker which is too short to allow pairing between the two domains on the same chain. Accordingly, the V_H and V_L domains of one fragment are forced to pair with the complementary V_L and V_H domains of another fragment, thereby forming two antigen-binding sites. Another strategy for making bispecific antibody fragments by the use of single-chain Fv (sFv) dimers has also been reported. See, Gruber et al., J. Immunol. 152:5368 (1994).

Antibodies with more than two valencies are contemplated. For example, trispecific antibodies can be prepared. Tutt et al., J. Immunol. 147:60 (1991). Exemplary bispecific antibodies can bind to two different epitopes, at least one of which originates in the protein antigen ofthe invention. Alternatively, an anti-antigenic arm of an immunoglobulin molecule can be combined with an arm which binds to a triggering molecule on a leukocyte such as a T-cell receptor molecule (e.g. CD2, CD3, CD28, or B7), or Fc receptors for IgG (FcγR), such as FcγRI (CD64), FcγRII (CD32) and FcγRIII (CD 16) so as to focus cellular defense mechanisms to the cell expressing the particular antigen. Bispecific antibodies can also be used to direct cytotoxic agents to cells which express a particular antigen. These antibodies possess an antigen-binding arm and an arm which binds a cytotoxic agent or a radionuclide chelator, such as EOTUBE, DPT A, DOTA, or TETA. Another bispecific antibody of interest binds the protein antigen described herein and further binds tissue factor (TF).

Heterocon jugate Antibodies

Heteroconjugate antibodies are also within the scope ofthe present invention. Heteroconjugate antibodies are composed of two covalently joined antibodies. Such antibodies have, for example, been proposed to target immune system cells to unwanted cells (U.S. Patent No. 4,676,980), and for treatment of HIV infection (WO 91/00360; WO 92/200373; EP 03089). It is contemplated that the antibodies can be prepared in vitro using known methods in synthetic protein chemistry, including those involving crosslinking agents. For example, immunotoxins can be constructed using a disulfide exchange reaction or by forming a thioether bond. Examples of suitable reagents for this purpose include iminothiolate and methyl-4-mercaptobutyrimidate and those disclosed, for example, in U.S. Patent No. 4,676,980.

Effector Function Engineering It can be desirable to modify the antibody of the invention with respect to effector function, so as to enhance, e.g., the effectiveness ofthe antibody in treating cancer. For example, cysteine residue(s) can be introduced into the Fc region, thereby allowing interchain disulfide bond formation in this region. The homodimeric antibody thus generated can have improved intemalization capability and/or increased complement-mediated cell killing and antibody-dependent cellular cytotoxicity (ADCC). See Caron et al., J. Exp Med., 176: 1191- 1195 (1992) and Shopes, J. Immunol., 148: 2918-2922 (1992). Homodimeric antibodies with enhanced anti-tumor activity can also be prepared using heterobifunctional cross-linkers as described in Wolff et al. Cancer Research, 53: 2560-2565 (1993). Alternatively, an antibody can be engineered that has dual Fc regions and can thereby have enhanced complement lysis and ADCC capabilities. See Stevenson et al., Anti-Cancer Drug Design, 3: 219-230 (1989).

Immunoconjugates

The invention also pertains to immunoconjugates comprising an antibody conjugated to a cytotoxic agent such as a chemotherapeutic agent, toxin (e.g., an enzymatically active toxin of bacterial, fungal, plant, or animal origin, or fragments thereof), or a radioactive isotope (i.e., a radioconjugate).

Chemotherapeutic agents useful in the generation of such immunoconjugates have been described above. Enzymatically active toxins and fragments thereof that can be used include diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. A variety of radionuclides are available for the production of radioconjugated antibodies. Examples include ²¹²Bi, ¹³¹I, ^{13 ,}In, ⁹⁰Y, and ¹⁸⁶Re.

Conjugates ofthe antibody and cytotoxic agent are made using a variety of bifunctional protein-coupling agents such as N-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane (IT), bifunctional derivatives of imidoesters (such as dimethyl adipimidate HCL), active esters (such as disuccinimidyl suberate), aldehydes (such as glutareldehyde), bis-azido compounds (such as bis (p-azidobenzoyl) hexanediamine), bis- diazonium derivatives (such as bis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such as tolyene 2,6-diisocyanate), and bis-active fluorine compounds (such as 1,5-difluoro- 2,4-dinitrobenzene). For example, a ricin immunotoxin can be prepared as described in Vitetta et al., Science, 238: 1098 (1987). Carbon- 14-labeled l-isothiocyanatobenzyl-3- methyldiethylene triaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent for conjugation of radionucleotide to the antibody. See WO94/11026.

In another embodiment, the antibody can be conjugated to a "receptor" (such streptavidin) for utilization in tumor pretargeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a "ligand" (e.g., avidin) that is in turn conjugated to a cytotoxic agent.

In one embodiment, methods for the screening of antibodies that possess the desired specificity include, but are not limited to, enzyme-linked immunosorbent assay (ELISA) and other immunologically-mediated techniques known within the art. In a specific embodiment, selection of antibodies that are specific to a particular domain of an NOVX protein is facilitated by generation of hybridomas that bind to the fragment of an NOVX protein possessing such a domain. Thus, antibodies that are specific for a desired domain within an NOVX protein, or derivatives, fragments, analogs or homologs thereof, are also provided herein.

Anti-NOVX antibodies may be used in methods known within the art relating to the localization and/or quantitation of an NOVX protein (e.g., for use in measuring levels ofthe NOVX protein within appropriate physiological samples, for use in diagnostic methods, for use in imaging the protein, and the like). In a given embodiment, antibodies for NOVX proteins, or derivatives, fragments, analogs or homologs thereof, that contain the antibody derived binding domain, are utilized as pharmacologically-active compounds (hereinafter "Therapeutics").

An anti-NOVX antibody (e.g., monoclonal antibody) can be used to isolate an NOVX polypeptide by standard techniques, such as affinity chromatography or immunoprecipitation. An anti-NOVX antibody can facilitate the purification of natural NOVX polypeptide from cells and of recombinantly-produced NOVX polypeptide expressed in host cells. Moreover, an anti-NOVX antibody can be used to detect NOVX protein (e.g., in a cellular lysate or cell supernatant) in order to evaluate the abundance and pattern of expression ofthe NOVX protein. Anti-NOVX antibodies can be used diagnostically to monitor protein levels in tissue as part of a clinical testing procedure, e.g., to, for example, determine the efficacy of a given treatment regimen. Detection can be facilitated by coupling (t.e., physically linking) the antibody to a detectable substance. Examples of detectable substances include various enzymes, prosthetic groups, fluorescent materials, luminescent materials, bioluminescent materials, and radioactive materials. Examples of suitable enzymes include horseradish peroxidase, alkaline phosphatase, β-galactosidase, or acetylcholinesterase; examples of suitable prosthetic group complexes include streptavidin/biotin and avidin/biotin; examples of suitable fluorescent materials include umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride or phycoerythrin; an example of a luminescent material includes luminol; examples of bioluminescent materials include luciferase, luciferin, and aequorin, and examples of suitable radioactive material include ¹²⁵I, ^!31I, ³⁵S or ³H.

NOVX Recombinant Expression Vectors and Host Cells

Another aspect ofthe invention pertains to vectors, preferably expression vectors, containing a nucleic acid encoding an NOVX protein, or derivatives, fragments, analogs or homologs thereof. As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non-episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. Moreover, certain vectors are capable of directing the expression of genes to which they are operatively-linked. Such vectors are referred to herein as "expression vectors". In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" can be used interchangeably as the plasmid is the most commonly used form of vector. However, the invention is intended to include such other forms of expression vectors, such as viral vectors (e.g. , replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions. The recombinant expression vectors ofthe invention comprise a nucleic acid ofthe invention in a form suitable for expression ofthe nucleic acid in a host cell, which means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis ofthe host cells to be used for expression, that is operatively-linked to the nucleic acid sequence to be expressed. Within a recombinant expression vector, "operably-linked" is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner that allows for expression ofthe nucleotide sequence (e.g., in an in vitro transcription translation system or in a host cell when the vector is introduced into the host cell).

The term "regulatory sequence" is intended to includes promoters, enhancers and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Regulatory sequences include those that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression ofthe nucleotide sequence only in certain host cells (e.g., tissue-specific regulatory sequences). It will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc. The expression vectors ofthe invention can be introduced into host cells to thereby produce proteins or peptides, including fusion proteins or peptides, encoded by nucleic acids as described herein (e.g., NOVX proteins, mutant forms of NOVX proteins, fusion proteins, etc.).

The recombinant expression vectors ofthe invention can be designed for expression of NOVX proteins in prokaryotic or eukaryotic cells. For example, NOVX proteins can be expressed in bacterial cells such as Escherichia coli, insect cells (using baculovirus expression vectors) yeast cells or mammalian cells. Suitable host cells are discussed further in Goeddel, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990). Alternatively, the recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase.

Expression of proteins in prokaryotes is most often carried out in Escherichia coli with vectors containing constitutive or inducible promoters directing the expression of either fusion or non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus ofthe recombinant protein. Such fusion vectors typically serve three purposes: (i) to increase expression of recombinant protein; (ii) to increase the solubility ofthe recombinant protein; and (ttt) to aid in the purification ofthe recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction ofthe fusion moiety and the recombinant protein to enable separation ofthe recombinant protein from the fusion moiety subsequent to purification ofthe fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, thrombin and enterokinase. Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith and Johnson, 1988. Gene 67: 31-40), pMAL (New England Biolabs, Beverly, Mass.) and pRIT5 (Pharmacia, Piscataway, NJ.) that fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein.

Examples of suitable inducible non- fusion E. coli expression vectors include pTrc (Amrann et al, (1988) Gene 69:301-315) and pET l id (Studier et al, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 60-89).

One strategy to maximize recombinant protein expression in E. coli is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein. See, e.g., Gottesman, GENE EXPRESSION TECHNOLOGY: METHODS IN ENZYMOLOGY 185, Academic Press, San Diego, Calif. (1990) 119-128. Another strategy is to alter the nucleic acid sequence ofthe nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in E. coli (see, e.g., Wada, et al, 1992. Nucl. Acids Res. 20: 2111-2118). Such alteration of nucleic acid sequences ofthe invention can be carried out by standard DNA synthesis techniques. In another embodiment, the NOVX expression vector is a yeast expression vector.

Examples of vectors for expression in yeast Saccharomyces cerivisae include pYepSecl (Baldari, et al, 1987. EMBO J. 6: 229-234), pMFa (Kurjan and Herskowitz, 1982. Cell 30: 933-943), pJRY88 (Schultz et al, 1987. Gene 54: 113-123), pYES2 (Invitrogen Corporation, San Diego, Calif), and picZ (InVitrogen Corp, San Diego, Calif.). Alternatively, NOVX can be expressed in insect cells using baculovirus expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., SF9 cells) include the pAc series (Smith, et al, 1983. Mol. Cell. Biol. 3: 2156-2165) and the pVL series (Lucklow and Summers, 1989. Virology 170: 31-39). In yet another embodiment, a nucleic acid ofthe invention is expressed in mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 (Seed, 1987. Nature 329: 840) and pMT2PC (Kaufman, et al, 1987. EMBO J. 6: 187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, adenovirus 2, cytomegalovirus, and simian vims 40. For other suitable expression systems for both prokaryotic and eukaryotic cells see, e.g., Chapters 16 and 17 of Sambrook, et al, MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989. In another embodiment, the recombinant mammalian expression vector is capable of directing expression of the nucleic acid preferentially in a particular cell type (e.g. , tissue-specific regulatory elements are used to express the nucleic acid). Tissue-specific regulatory elements are known in the art. Non- limiting examples of suitable tissue-specific promoters include the albumin promoter (liver-specific; Pinkert, et al, 1987. Genes Dev. 1 : 268-277), lymphoid-specific promoters (Calame and Eaton, 1988. Adv. Immunol. 43: 235-275), in particular promoters of T cell receptors (Winoto and Baltimore, 1989. EMBO J. 8: 729-733) and immunoglobulins (Banerji, et al, 1983. Cell 33: 729-740; Queen and Baltimore, 1983. Cell 33: 741-748), neuron-specific promoters (e.g., the neurofilament promoter; Byrne and Ruddle, 1989. Proc. Natl. Acad. Sci. USA 86: 5473-5477), pancreas-specific promoters (Edlund, et al, 1985. Science 230: 912-916), and mammary gland-specific promoters (e.g. , milk whey promoter; U.S. Pat. No. 4,873,316 and European Application Publication No. 264,166). Developmentally-regulated promoters are also encompassed, e.g., the murine hox promoters (Kessel and Gruss, 1990. Science 249: 374-379) and the α-fetoprotein promoter (Campes and Tilghman, 1989. Genes Dev. 3: 537-546).

The invention further provides a recombinant expression vector comprising a DNA molecule ofthe invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively-linked to a regulatory sequence in a manner that allows for expression (by transcription ofthe DNA molecule) of an RNA molecule that is antisense to NOVX mRNA. Regulatory sequences operatively linked to a nucleic acid cloned in the antisense orientation can be chosen that direct the continuous expression ofthe antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen that direct constitutive, tissue specific or cell type specific expression of antisense RNA. The antisense expression vector can be in the form of a recombinant plasmid, phagemid or attenuated vims in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced. For a discussion ofthe regulation of gene expression using antisense genes see, e.g., Weintraub, et al, "Antisense RNA as a molecular tool for genetic analysis," Reviews-Trends in Genetics, Vol. 1(1) 1986.

Another aspect ofthe invention pertains to host cells into which a recombinant expression vector ofthe invention has been introduced. The terms "host cell" and

"recombinant host cell" are used interchangeably herein. It is understood that such terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope ofthe term as used herein.

A host cell can be any prokaryotic or eukaryotic cell. For example, NOVX protein can be expressed in bacterial cells such as E. coli, insect cells, yeast or mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells). Other suitable host cells are known to those skilled in the art. Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms "transformation" and "transfection" are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DΕAΕ-dextran-mediated transfection, lipofection, or electroporation. Suitable methods for transforming or transfecting host cells can be found in Sambrook, et al. (MOLECULAR CLONING: A LABORATORY MANUAL. 2nd ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., 1989), and other laboratory manuals.

For stable transfection of mammalian cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to antibiotics) is generally introduced into the host cells along with the gene of interest. Various selectable markers include those that confer resistance to drugs, such as G418, hygromycin and methotrexate. Nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding NOVX or can be introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by dmg selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die). A host cell ofthe invention, such as a prokaryotic or eukaryotic host cell in culture, can be used to produce (t.e., express) NOVX protein. Accordingly, the invention further provides methods for producing NOVX protein using the host cells ofthe invention. In one embodiment, the method comprises culturing the host cell of invention (into which a recombinant expression vector encoding NOVX protein has been introduced) in a suitable medium such that NOVX protein is produced. In another embodiment, the method further comprises isolating NOVX protein from the medium or the host cell.

Transgenic NOVX Animals

The host cells ofthe invention can also be used to produce non-human transgenic animals. For example, in one embodiment, a host cell ofthe invention is a fertilized oocyte or an embryonic stem cell into which NOVX protein-coding sequences have been introduced. Such host cells can then be used to create non-human transgenic animals in which exogenous NOVX sequences have been introduced into their genome or homologous recombinant animals in which endogenous NOVX sequences have been altered. Such animals are useful for studying the function and/or activity of NOVX protein and for identifying and/or evaluating modulators of NOVX protein activity. As used herein, a "transgenic animal" is a non-human animal, preferably a mammal, more preferably a rodent such as a rat or mouse, in which one or more ofthe cells ofthe animal includes a transgene. Other examples of transgenic animals include non-human primates, sheep, dogs, cows, goats, chickens, amphibians, etc. A transgene is exogenous DNA that is integrated into the genome of a cell from which a transgenic animal develops and that remains in the genome ofthe mature animal, thereby directing the expression of an encoded gene product in one or more cell types or tissues ofthe transgenic animal. As used herein, a "homologous recombinant animal" is a non-human animal, preferably a mammal, more preferably a mouse, in which an endogenous NOVX gene has been altered by homologous recombination between the endogenous gene and an exogenous DNA molecule introduced into a cell ofthe animal, e.g., an embryonic cell ofthe animal, prior to development ofthe animal.

A transgenic animal ofthe invention can be created by introducing NOVX-encoding nucleic acid into the male pronuclei of a fertilized oocyte (e.g., by microinjection, retroviral infection) and allowing the oocyte to develop in a pseudopregnant female foster animal. The human NOVX cDNA sequences SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 can be introduced as a transgene into the genome of a non-human animal. Alternatively, a non-human homologue ofthe human NOVX gene, such as a mouse NOVX gene, can be isolated based on hybridization to the human NOVX cDNA (described further supra) and used as a transgene. Intronic sequences and polyadenylation signals can also be included in the transgene to increase the efficiency of expression ofthe transgene. A tissue-specific regulatory sequence(s) can be operably-linked to the NOVX transgene to direct expression of NOVX protein to particular cells. Methods for generating transgenic animals via embryo manipulation and microinjection, particularly animals such as mice, have become conventional in the art and are described, for example, in U.S. Patent Nos. 4,736,866; 4,870,009; and 4,873,191; and Hogan, 1986. In: MANIPULATING THE MOUSE EMBRYO, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Similar methods are used for production of other transgenic animals. A transgenic founder animal can be identified based upon the presence ofthe NOVX transgene in its genome and/or expression of NOVX mRNA in tissues or cells ofthe animals. A transgenic founder animal can then be used to breed additional animals carrying the transgene. Moreover, transgenic animals carrying a transgene-encoding NOVX protein can further be bred to other transgenic animals carrying other transgenes.

To create a homologous recombinant animal, a vector is prepared which contains at least a portion of an NOVX gene into which a deletion, addition or substitution has been introduced to thereby alter, e.g., functionally disrupt, the NOVX gene. The NOVX gene can be a human gene (e.g., the cDNA of SEQ LD NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47), but more preferably, is a non-human homologue of a human NOVX gene. For example, a mouse homologue of human NOVX gene of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 can be used to constmct a homologous recombination vector suitable for altering an endogenous NOVX gene in the mouse genome. In one embodiment, the vector is designed such that, upon homologous recombination, the endogenous NOVX gene is functionally disrupted (t.e., no longer encodes a functional protein; also referred to as a "knock out" vector).

Alternatively, the vector can be designed such that, upon homologous recombination, the endogenous NOVX gene is mutated or otherwise altered but still encodes functional protein (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous NOVX protein). In the homologous recombination vector, the altered portion ofthe NOVX gene is flanked at its 5'- and 3'-termini by additional nucleic acid ofthe NOVX gene to allow for homologous recombination to occur between the exogenous NOVX gene carried by the vector and an endogenous NOVX gene in an embryonic stem cell. The additional flanking NOVX nucleic acid is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several kilobases of flanking DNA (both at the 5'- and 3'-termini) are included in the vector. See, e.g., Thomas, et al, 1987. Cell 51 : 503 for a description of homologous recombination vectors. The vector is ten introduced into an embryonic stem cell line (e.g., by electroporation) and cells in which the introduced NOVX gene has homologously-recombined with the endogenous NOVX gene are selected. See, e.g., U, et al, 1992. Cell 69: 915.

The selected cells are then injected into a blastocyst of an animal (e.g., a mouse) to form aggregation chimeras. See, e.g., Bradley, 1987. In: TERATOCARCINOMAS AND EMBRYONIC STEM CELLS: A PRACTICAL APPROACH, Robertson, ed. LRL, Oxford, pp. 113-152. A chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal and the embryo brought to term. Progeny harboring the homologously-recombined DNA in their germ cells can be used to breed animals in which all cells ofthe animal contain the homologously-recombined DNA by germline transmission ofthe transgene. Methods for constmcting homologous recombination vectors and homologous recombinant animals are described further in Bradley, 1991. Curr. Opin. Biotechnol. 2: 823-829; PCT International Publication Nos.: WO 90/11354; WO 91/01140; WO 92/0968; and WO 93/04169.

In another embodiment, transgenic non-humans animals can be produced that contain selected systems that allow for regulated expression ofthe transgene. One example of such a system is the cre/loxP recombinase system of bacteriophage PI. For a description ofthe cre/loxP recombinase system, See, e.g., Lakso, et al, 1992. Proc. Natl. Acad. Sci. USA 89: 6232-6236. Another example of a recombinase system is the FLP recombinase system of

Saccharomyces cerevisiae. See, O'Gorman, et al, 1991. Science 251:1351-1355. Ifa cre/loxP recombinase system is used to regulate expression ofthe transgene, animals containing transgenes encoding both the Cre recombinase and a selected protein are required. Such animals can be provided through the construction of "double" transgenic animals, e.g., by mating two transgenic animals, one containing a transgene encoding a selected protein and the other containing a transgene encoding a recombinase.

Clones ofthe non-human transgenic animals described herein can also be produced according to the methods described in Wilmut, et al, 1997. Nature 385: 810-813. In brief, a cell (e.g., a somatic cell) from the transgenic animal can be isolated and induced to exit the growth cycle and enter G₀ phase. The quiescent cell can then be fused, e.g., through the use of electrical pulses, to an enucleated oocyte from an animal ofthe same species from which the quiescent cell is isolated. The reconstmcted oocyte is then cultured such that it develops to momla or blastocyte and then transferred to pseudopregnant female foster animal. The offspring borne of this female foster animal will be a clone ofthe animal from which the cell (e.g., the somatic cell) is isolated.

Pharmaceutical Compositions

The NOVX nucleic acid molecules, NOVX proteins, and anti-NOVX antibodies (also referred to herein as "active compounds") ofthe invention, and derivatives, fragments, analogs and homologs thereof, can be incorporated into pharmaceutical compositions suitable for administration. Such compositions typically comprise the nucleic acid molecule, protein, or antibody and a pharmaceutically acceptable carrier. As used herein, "pharmaceutically acceptable carrier" is intended to include any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, compatible with pharmaceutical administration. Suitable carriers are described in the most recent edition of Remington's Pharmaceutical Sciences, a standard reference text in the field, which is incorporated herein by reference. Preferred examples of such carriers or diluents include, but are not limited to, water, saline, finger's solutions, dextrose solution, and 5% human serum albumin. Liposomes and non-aqueous vehicles such as fixed oils may also be used. The use of such media and agents for pharmaceutically active substances is well known in the art. Except insofar as any conventional media or agent is incompatible with the active compound, use thereof in the compositions is contemplated. Supplementary active compounds can also be incorporated into the compositions.

A pharmaceutical composition ofthe invention is formulated to be compatible with its intended route of administration. Examples of routes of administration include parenteral, e.g., intravenous, intradermal, subcutaneous, oral (e.g., inhalation), transdermal (t.e., topical), transmucosal, and rectal administration. Solutions or suspensions used for parenteral, intradermal, or subcutaneous application can include the following components: a sterile diluent such as water for injection, saline solution, fixed oils, polyethylene glycols, glycerine, propylene glycol or other synthetic solvents; antibacterial agents such as benzyl alcohol or methyl parabens; antioxidants such as ascorbic acid or sodium bisulfite; chelating agents such as ethylenediaminetetraacetic acid (EDTA); buffers such as acetates, citrates or phosphates, and agents for the adjustment of tonicity such as sodium chloride or dextrose. The pH can be adjusted with acids or bases, such as hydrochloric acid or sodium hydroxide. The parenteral preparation can be enclosed in ampoules, disposable syringes or multiple dose vials made of glass or plastic.

Pharmaceutical compositions suitable for injectable use include sterile aqueous solutions (where water soluble) or dispersions and sterile powders for the extemporaneous preparation of sterile injectable solutions or dispersion. For intravenous administration, suitable carriers include physiological saline, bacteriostatic water, Cremophor EL (BASF, Parsippany, NJ.) or phosphate buffered saline (PBS). In all cases, the composition must be sterile and should be fluid to the extent that easy syringeability exists. It must be stable under the conditions of manufacture and storage and must be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by the maintenance ofthe required particle size in the case of dispersion and by the use of surfactants. Prevention ofthe action of microorganisms can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, ascorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, polyalcohols such as manitol, sorbitol, sodium chloride in the composition. Prolonged absorption ofthe injectable compositions can be brought about by including in the composition an agent which delays absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions can be prepared by incorporating the active compound (e.g., an NOVX protein or anti-NOVX antibody) in the required amount in an appropriate solvent with one or a combination of ingredients enumerated above, as required, followed by filtered sterilization. Generally, dispersions are prepared by incorporating the active compound into a sterile vehicle that contains a basic dispersion medium and the required other ingredients from those enumerated above. In the case of sterile powders for the preparation of sterile injectable solutions, methods of preparation are vacuum drying and freeze-drying that yields a powder of the active ingredient plus any additional desired ingredient from a previously sterile-filtered solution thereof.

Oral compositions generally include an inert diluent or an edible carrier. They can be enclosed in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, troches, or capsules. Oral compositions can also be prepared using a fluid carrier for use as a mouthwash, wherein the compound in the fluid carrier is applied orally and swished and expectorated or swallowed. Pharmaceutically compatible binding agents, and/or adjuvant materials can be included as part ofthe composition. The tablets, pills, capsules, troches and the like can contain any ofthe following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, gum tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel, or com starch; a lubricant such as magnesium stearate or Sterotes; a glidant such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin; or a flavoring agent such as peppermint, methyl salicylate, or orange flavoring. For administration by inhalation, the compounds are delivered in the form of an aerosol spray from pressured container or dispenser which contains a suitable propellant, e.g., a gas such as carbon dioxide, or a nebulizer.

Systemic administration can also be by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art, and include, for example, for transmucosal administration, detergents, bile salts, and fusidic acid derivatives. Transmucosal administration can be accomplished through the use of nasal sprays or suppositories. For transdermal administration, the active compounds are formulated into ointments, salves, gels, or creams as generally known in the art. The compounds can also be prepared in the form of suppositories (e.g. , with conventional suppository bases such as cocoa butter and other glycerides) or retention enemas for rectal delivery.

In one embodiment, the active compounds are prepared with carriers that will protect the compound against rapid elimination from the body, such as a controlled release formulation, including implants and microencapsulated delivery systems. Biodegradable, biocompatible polymers can be used, such as ethylene vinyl acetate, polyanhydrides, polyglycolic acid, collagen, polyorthoesters, and polylactic acid. Methods for preparation of such formulations will be apparent to those skilled in the art. The materials can also be obtained commercially from Alza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions (including liposomes targeted to infected cells with monoclonal antibodies to viral antigens) can also be used as pharmaceutically acceptable carriers. These can be prepared according to methods known to those skilled in the art, for example, as described in U.S. Patent No. 4,522,811. It is especially advantageous to formulate oral or parenteral compositions in dosage unit form for ease of administration and uniformity of dosage. Dosage unit form as used herein refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on the unique characteristics ofthe active compound and the particular therapeutic effect to be achieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals. The nucleic acid molecules ofthe invention can be inserted into vectors and used as gene therapy vectors. Gene therapy vectors can be delivered to a subject by, for example, intravenous injection, local administration (see, e.g., U.S. Patent No. 5,328,470) or by stereotactic injection (see, e.g., Chen, et al, 1994. Proc. Natl. Acad. Sci. USA 91 : 3054-3057). The pharmaceutical preparation ofthe gene therapy vector can include the gene therapy vector in an acceptable diluent, or can comprise a slow release matrix in which the gene delivery vehicle is imbedded. Alternatively, where the complete gene delivery vector can be produced intact from recombinant cells, e.g., retroviral vectors, the pharmaceutical preparation can include one or more cells that produce the gene delivery system.

The pharmaceutical compositions can be included in a container, pack, or dispenser together with instmctions for administration.

Screening and Detection Methods

The isolated nucleic acid molecules ofthe invention can be used to express NOVX protein (e.g., via a recombinant expression vector in a host cell in gene therapy applications), to detect NOVX mRNA (e.g. , in a biological sample) or a genetic lesion in an NOVX gene, and to modulate NOVX activity, as described further, below. In addition, the NOVX proteins can be used to screen drugs or compounds that modulate the NOVX protein activity or expression as well as to treat disorders characterized by insufficient or excessive production of NOVX protein or production of NOVX protein forms that have decreased or aberrant activity compared to NOVX wild-type protein (e.g.; diabetes (regulates insulin release); obesity (binds and transport lipids); metabolic disturbances associated with obesity, the metabolic syndrome X as well as anorexia and wasting disorders associated with chronic diseases and various cancers, and infectious disease(possesses anti-microbial activity) and the various dyslipidemias. In addition, the anti-NOVX antibodies ofthe invention can be used to detect and isolate NOVX proteins and modulate NOVX activity. In yet a further aspect, the invention can be used in methods to influence appetite, absorption of nutrients and the disposition of metabolic substrates in both a positive and negative fashion.

The invention further pertains to novel agents identified by the screening assays described herein and uses thereof for treatments as described, supra. Screening Assays

The invention provides a method (also referred to herein as a "screening assay") for identifying modulators, i.e., candidate or test compounds or agents (e.g., peptides, peptidomimetics, small molecules or other drugs) that bind to NOVX proteins or have a stimulatory or inhibitory effect on, e.g., NOVX protein expression or NOVX protein activity. The invention also includes compounds identified in the screening assays described herein. In one embodiment, the invention provides assays for screening candidate or test compounds which bind to or modulate the activity ofthe membrane-bound form of an NOVX protein or polypeptide or biologically-active portion thereof. The test compounds ofthe invention can be obtained using any ofthe numerous approaches in combinatorial library methods known in the art, including: biological libraries; spatially addressable parallel solid phase or solution phase libraries; synthetic library methods requiring deconvolution; the "one-bead one-compound" library method; and synthetic library methods using affinity chromatography selection. The biological library approach is limited to peptide libraries, while the other four approaches are applicable to peptide, non-peptide oligomer or small molecule libraries of compounds. See, e.g., Lam, 199 '. Anticancer Drug Design 12: 145. A "small molecule" as used herein, is meant to refer to a composition that has a molecular weight of less than about 5 kD and most preferably less than about 4 kD. Small molecules can be, e.g., nucleic acids, peptides, polypeptides, peptidomimetics, carbohydrates, lipids or other organic or inorganic molecules. Libraries of chemical and/or biological mixtures, such as fungal, bacterial, or algal extracts, are known in the art and can be screened with any ofthe assays ofthe invention.

Examples of methods for the synthesis of molecular libraries can be found in the art, for example in: DeWitt, et al, 1993. Proc. Natl. Acad. Sci. U.S.A. 90: 6909; Erb, et al, 1994. Proc. Natl Acad. Sci. U.S.A. 91 : 11422; Zuckermann, et al, 1994. J. Med. Chem. 37: 2678; Cho, et al, 1993. Science 261 : 1303; Carrell, et al, 1994. Angew. Chem. Int. Ed. Engl. 33: 2059; Carell, et al, 1994. Angew. Chem. Int. Ed. Engl. 33: 2061; and Gallop, et al, 1994. J. Med. Chem. 37: 1233.

Libraries of compounds may be presented in solution (e.g., Houghten, 1992. Biotechniques 13: 412-421), or on beads (Lam, 1991. Nature 354: 82-84), on chips (Fodor, 1993. Nature 364: 555-556), bacteria (Ladner, U.S. Patent No. 5,223,409), spores (Ladner, U.S. Patent 5,233,409), plasmids (Cull, et al, 1992. Proc. Natl. Acad. Sci. USA 89: 1865-1869) or on phage (Scott and Smith, 1990. Science 249: 386-390; Devlin, 1990. Science 249: 404-406; Cwirla, et al, 1990. Proc. Natl. Acad. Sci. U.S.A. 87: 6378-6382; Felici, 1991. J. Mol. Biol. 222: 301-310; Ladner, U.S. Patent No. 5,233,409.).

In one embodiment, an assay is a cell-based assay in which a cell which expresses a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface is contacted with a test compound and the ability ofthe test compound to bind to an NOVX protein determined. The cell, for example, can of mammalian origin or a yeast cell. Determining the ability of the test compound to bind to the NOVX protein can be accomplished, for example, by coupling the test compound with a radioisotope or enzymatic label such that binding ofthe test compound to the NOVX protein or biologically-active portion thereof can be determined by detecting the labeled compound in a complex. For example, test compounds can be labeled with ¹²⁵1, ³⁵S, ¹⁴C, or ³H, either directly or indirectly, and the radioisotope detected by direct counting of radioemission or by scintillation counting. Alternatively, test compounds can be enzymatically-labeled with, for example, horseradish peroxidase, alkaline phosphatase, or luciferase, and the enzymatic label detected by determination of conversion of an appropriate substrate to product. In one embodiment, the assay comprises contacting a cell which expresses a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface with a known compound which binds NOVX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability ofthe test compound to interact with an NOVX protein, wherein determining the ability of the test compound to interact with an NOVX protein comprises determining the ability of the test compound to preferentially bind to NOVX protein or a biologically-active portion thereof as compared to the known compound.

In another embodiment, an assay is a cell-based assay comprising contacting a cell expressing a membrane-bound form of NOVX protein, or a biologically-active portion thereof, on the cell surface with a test compound and determining the ability ofthe test compound to modulate (e.g., stimulate or inhibit) the activity ofthe NOVX protein or biologically-active portion thereof. Determining the ability ofthe test compound to modulate the activity of NOVX or a biologically-active portion thereof can be accomplished, for example, by determining the ability ofthe NOVX protein to bind to or interact with an NOVX target molecule. As used herein, a "target molecule" is a molecule with which an NOVX protein binds or interacts in nature, for example, a molecule on the surface of a cell which expresses an NOVX interacting protein, a molecule on the surface of a second cell, a molecule in the extracellular milieu, a molecule associated with the internal surface of a cell membrane or a cytoplasmic molecule. An NOVX target molecule can be a non-NOVX molecule or an NOVX protein or polypeptide ofthe invention. In one embodiment, an NOVX target molecule is a component of a signal transduction pathway that facilitates transduction of an extracellular signal (e.g. a signal generated by binding of a compound to a membrane-bound NOVX molecule) through the cell membrane and into the cell. The target, for example, can be a second intercellular protein that has catalytic activity or a protein that facilitates the association of downstream signaling molecules with NOVX. Determining the ability ofthe NOVX protein to bind to or interact with an NOVX target molecule can be accomplished by one ofthe methods described above for determining direct binding. In one embodiment, determining the ability ofthe NOVX protein to bind to or interact with an NOVX target molecule can be accomplished by determining the activity ofthe target molecule. For example, the activity ofthe target molecule can be determined by detecting induction of a cellular second messenger ofthe target (t.e. intracellular Ca²⁺, diacylglycerol, IP₃, etc.), detecting catalytic/enzymatic activity ofthe target an appropriate substrate, detecting the induction of a reporter gene (comprising an NOVX-responsive regulatory element operatively linked to a nucleic acid encoding a detectable marker, e.g., luciferase), or detecting a cellular response, for example, cell survival, cellular differentiation, or cell proliferation.

In yet another embodiment, an assay ofthe invention is a cell- free assay comprising contacting an NOVX protein or biologically-active portion thereof with a test compound and determining the ability ofthe test compound to bind to the NOVX protein or biologically- active portion thereof. Binding ofthe test compound to the NOVX protein can be determined either directly or indirectly as described above. In one such embodiment, the assay comprises contacting the NOVX protein or biologically-active portion thereof with a known compound which binds NOVX to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability ofthe test compound to interact with an NOVX protein, wherein determining the ability ofthe test compound to interact with an NOVX protein comprises determining the ability ofthe test compound to preferentially bind to NOVX or biologically-active portion thereof as compared to the known compound.

In still another embodiment, an assay is a cell-free assay comprising contacting NOVX protein or biologically-active portion thereof with a test compound and determining the ability ofthe test compound to modulate (e.g. stimulate or inhibit) the activity of the NOVX protein or biologically-active portion thereof. Determining the ability ofthe test compound to modulate the activity of NOVX can be accomplished, for example, by determining the ability ofthe NOVX protein to bind to an NOVX target molecule by one ofthe methods described above for determining direct binding. In an alternative embodiment, determining the ability of the test compound to modulate the activity of NOVX protein can be accomplished by determining the ability ofthe NOVX protein further modulate an NOVX target molecule. For example, the catalytic/enzymatic activity ofthe target molecule on an appropriate substrate can be determined as described, supra.

In yet another embodiment, the cell-free assay comprises contacting the NOVX protein or biologically-active portion thereof with a known compound which binds NOVX protein to form an assay mixture, contacting the assay mixture with a test compound, and determining the ability ofthe test compound to interact with an NOVX protein, wherein determining the ability ofthe test compound to interact with an NOVX protein comprises determining the ability ofthe NOVX protein to preferentially bind to or modulate the activity of an NOVX target molecule.

The cell- free assays ofthe invention are amenable to use of both the soluble form or the membrane-bound form of NOVX protein. In the case of cell-free assays comprising the membrane-bound form of NOVX protein, it may be desirable to utilize a solubilizing agent such that the membrane-bound form of NOVX protein is maintained in solution. Examples of such solubilizing agents include non-ionic detergents such as n-octylglucoside, n-dodecylglucoside, n-dodecylmaltoside, octanoyl-N-methylglucamide, decanoyl-N-methylglucamide, Triton^® X-100, Triton^® X-114, Thesit^®, Isotridecypoly(ethylene glycol ether)_n, N-dodecyl~N,N-dimethyl-3-ammonio-l -propane sulfonate, 3-(3-cholamidopropyl) dimethylamminiol-1 -propane sulfonate (CHAPS), or 3-(3-cholamidopropyl)dimethylamminiol-2-hydroxy-l -propane sulfonate (CHAPSO).

In more than one embodiment ofthe above assay methods ofthe invention, it may be desirable to immobilize either NOVX protein or its target molecule to facilitate separation of complexed from uncomplexed forms of one or both ofthe proteins, as well as to accommodate automation ofthe assay. Binding of a test compound to NOVX protein, or interaction of NOVX protein with a target molecule in the presence and absence of a candidate compound, can be accomplished in any vessel suitable for containing the reactants. Examples of such vessels include microtiter plates, test tubes, and micro-centrifuge tubes. In one embodiment, a fusion protein can be provided that adds a domain that allows one or both ofthe proteins to be bound to a matrix. For example, GST-NOVX fusion proteins or GST-target fusion proteins can be adsorbed onto glutathione sepharose beads (Sigma Chemical, St. Louis, MO) or glutathione derivatized microtiter plates, that are then combined with the test compound or the test compound and either the non-adsorbed target protein or NOVX protein, and the mixture is incubated under conditions conducive to complex formation (e.g., at physiological conditions for salt and pH). Following incubation, the beads or microtiter plate wells are washed to remove any unbound components, the matrix immobilized in the case of beads, complex determined either directly or indirectly, for example, as described, supra. Alternatively, the complexes can be dissociated from the matrix, and the level of NOVX protein binding or activity determined using standard techniques. Other techniques for immobilizing proteins on matrices can also be used in the screening assays ofthe invention. For example, either the NOVX protein or its target molecule can be immobilized utilizing conjugation of biotin and streptavidin. Biotinylated NOVX protein or target molecules can be prepared from biotin-NHS (N-hydroxy-succinimide) using techniques well-known within the art (e.g., biotinylation kit, Pierce Chemicals, Rockford, 111.), and immobilized in the wells of streptavidin-coated 96 well plates (Pierce Chemical). Alternatively, antibodies reactive with NOVX protein or target molecules, but which do not interfere with binding ofthe NOVX protein to its target molecule, can be derivatized to the wells ofthe plate, and unbound target or NOVX protein trapped in the wells by antibody conjugation. Methods for detecting such complexes, in addition to those described above for the GST-immobilized complexes, include immunodetection of complexes using antibodies reactive with the NOVX protein or target molecule, as well as enzyme-linked assays that rely on detecting an enzymatic activity associated with the NOVX protein or target molecule.

In another embodiment, modulators of NOVX protein expression are identified in a method wherein a cell is contacted with a candidate compound and the expression of NOVX mRNA or protein in the cell is determined. The level of expression of NOVX mRNA or protein in the presence ofthe candidate compound is compared to the level of expression of NOVX mRNA or protein in the absence ofthe candidate compound. The candidate compound can then be identified as a modulator of NOVX mRNA or protein expression based upon this comparison. For example, when expression of NOVX mRNA or protein is greater (t.e., statistically significantly greater) in the presence ofthe candidate compound than in its absence, the candidate compound is identified as a stimulator of NOVX mRNA or protein expression. Alternatively, when expression of NOVX mRNA or protein is less (statistically significantly less) in the presence ofthe candidate compound than in its absence, the candidate compound is identified as an inhibitor of NOVX mRNA or protein expression. The level of NOVX mRNA or protein expression in the cells can be determined by methods described herein for detecting NOVX mRNA or protein.

In yet another aspect ofthe invention, the NOVX proteins can be used as "bait proteins" in a two-hybrid assay or three hybrid assay (see, e.g., U.S. Patent No. 5,283,317;

Zervos, et al, 1993. Cell 72: 223-232; Madura, et al, 1993. J. Biol Chem. 268: 12046-12054; Bartel, et al, 1993. Biotechniques 14: 920-924; Iwabuchi, et al, 1993. Oncogene 8: 1693-1696; and Brent WO 94/10300), to identify other proteins that bind to or interact with NOVX ("NOVX-binding proteins" or "NOVX-bp") and modulate NOVX activity. Such NOVX-binding proteins are also likely to be involved in the propagation of signals by the NOVX proteins as, for example, upstream or downstream elements ofthe NOVX pathway. The two-hybrid system is based on the modular nature of most transcription factors, which consist of separable DNA-binding and activation domains. Briefly, the assay utilizes two different DNA constructs. In one constmct, the gene that codes for NOVX is fused to a gene encoding the DNA binding domain of a known transcription factor (e.g. , GAL-4). In the other constmct, a DNA sequence, from a library of DNA sequences, that encodes an unidentified protein ("prey" or "sample") is fused to a gene that codes for the activation domain ofthe known transcription factor. If the "bait" and the "prey" proteins are able to interact, in vivo, forming an NOVX-dependent complex, the DNA-binding and activation domains ofthe transcription factor are brought into close proximity. This proximity allows transcription of a reporter gene (e.g., LacZ) that is operably linked to a transcriptional regulatory site responsive to the transcription factor. Expression ofthe reporter gene can be detected and cell colonies containing the functional transcription factor can be isolated and used to obtain the cloned gene that encodes the protein which interacts with NOVX. The invention further pertains to novel agents identified by the aforementioned screening assays and uses thereof for treatments as described herein.

Detection Assays

Portions or fragments ofthe cDNA sequences identified herein (and the corresponding complete gene sequences) can be used in numerous ways as polynucleotide reagents. By way of example, and not of limitation, these sequences can be used to: (i) map their respective genes on a chromosome; and, thus, locate gene regions associated with genetic disease; (ii) identify an individual from a minute biological sample (tissue typing); and (ttt) aid in forensic identification of a biological sample. Some of these applications are described in the subsections, below. Chromosome Mapping

Once the sequence (or a portion ofthe sequence) of a gene has been isolated, this sequence can be used to map the location ofthe gene on a chromosome. This process is called chromosome mapping. Accordingly, portions or fragments ofthe NOVX sequences, SEQ LD NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or fragments or derivatives thereof, can be used to map the location ofthe NOVX genes, respectively, on a chromosome. The mapping ofthe NOVX sequences to chromosomes is an important first step in correlating these sequences with genes associated with disease.

Briefly, NOVX genes can be mapped to chromosomes by preparing PCR primers (preferably 15-25 bp in length) from the NOVX sequences. Computer analysis ofthe NOVX, sequences can be used to rapidly select primers that do not span more than one exon in the genomic DNA, thus complicating the amplification process. These primers can then be used for PCR screening of somatic cell hybrids containing individual human chromosomes. Only those hybrids containing the human gene corresponding to the NOVX sequences will yield an amplified fragment.

Somatic cell hybrids are prepared by fusing somatic cells from different mammals (e.g., human and mouse cells). As hybrids of human and mouse cells grow and divide, they gradually lose human chromosomes in random order, but retain the mouse chromosomes. By using media in which mouse cells cannot grow, because they lack a particular enzyme, but in which human cells can, the one human chromosome that contains the gene encoding the needed enzyme will be retained. By using various media, panels of hybrid cell lines can be established. Each cell line in a panel contains either a single human chromosome or a small number of human chromosomes, and a full set of mouse chromosomes, allowing easy mapping of individual genes to specific human chromosomes. See, e.g., D'Eustachio, et al, 1983. Science 220: 919-924. Somatic cell hybrids containing only fragments of human chromosomes can also be produced by using human chromosomes with translocations and deletions.

PCR mapping of somatic cell hybrids is a rapid procedure for assigning a particular sequence to a particular chromosome. Three or more sequences can be assigned per day using a single thermal cycler. Using the NOVX sequences to design oligonucleotide primers, sub- localization can be achieved with panels of fragments from specific chromosomes.

Fluorescence in situ hybridization (FISH) of a DNA sequence to a metaphase chromosomal spread can further be used to provide a precise chromosomal location in one step. Chromosome spreads can be made using cells whose division has been blocked in metaphase by a chemical like colcemid that dismpts the mitotic spindle. The chromosomes can be treated briefly with trypsin, and then stained with Giemsa. A pattern of light and dark bands develops on each chromosome, so that the chromosomes can be identified individually. The FISH technique can be used with a DNA sequence as short as 500 or 600 bases. However, clones larger than 1,000 bases have a higher likelihood of binding to a unique chromosomal location with sufficient signal intensity for simple detection. Preferably 1 ,000 bases, and more preferably 2,000 bases, will suffice to get good results at a reasonable amount of time. For a review of this technique, see, Verma, et al, HUMAN CHROMOSOMES: A MANUAL OF BASIC TECHNIQUES (Pergamon Press, New York 1988). Reagents for chromosome mapping can be used individually to mark a single chromosome or a single site on that chromosome, or panels of reagents can be used for marking multiple sites and/or multiple chromosomes. Reagents corresponding to noncoding regions ofthe genes actually are preferred for mapping purposes. Coding sequences are more likely to be conserved within gene families, thus increasing the chance of cross hybridizations during chromosomal mapping.

Once a sequence has been mapped to a precise chromosomal location, the physical position ofthe sequence on the chromosome can be correlated with genetic map data. Such data are found, e.g., in McKusick, MENDELIAN INHERITANCE IN MAN, available on-line through Johns Hopkins University Welch Medical Library). The relationship between genes and disease, mapped to the same chromosomal region, can then be identified through linkage analysis (co-inheritance of physically adjacent genes), described in, e.g., Egeland, et al, 1987. Nature, 325: 783-787.

Moreover, differences in the DNA sequences between individuals affected and unaffected with a disease associated with the NOVX gene, can be determined. If a mutation is observed in some or all ofthe affected individuals but not in any unaffected individuals, then the mutation is likely to be the causative agent ofthe particular disease. Comparison of affected and unaffected individuals generally involves first looking for structural alterations in the chromosomes, such as deletions or translocations that are visible from chromosome spreads or detectable using PCR based on that DNA sequence. Ultimately, complete sequencing of genes from several individuals can be performed to confirm the presence of a mutation and to distinguish mutations from polymorphisms.

Tissue Typing

The NOVX sequences of the invention can also be used to identify individuals from minute biological samples. In this technique, an individual's genomic DNA is digested with one or more restriction enzymes, and probed on a Southern blot to yield unique bands for identification. The sequences ofthe invention are useful as additional DNA markers for RFLP ("restriction fragment length polymorphisms," described in U.S. Patent No. 5,272,057). Furthermore, the sequences ofthe invention can be used to provide an alternative technique that determines the actual base-by-base DNA sequence of selected portions of an individual's genome. Thus, the NOVX sequences described herein can be used to prepare two PCR primers from the 5'- and 3'-termini ofthe sequences. These primers can then be used to amplify an individual's DNA and subsequently sequence it.

Panels of corresponding DNA sequences from individuals, prepared in this manner, can provide unique individual identifications, as each individual will have a unique set of such DNA sequences due to allelic differences. The sequences ofthe invention can be used to obtain such identification sequences from individuals and from tissue. The NOVX sequences ofthe invention uniquely represent portions ofthe human genome. Allelic variation occurs to some degree in the coding regions of these sequences, and to a greater degree in the noncoding regions. It is estimated that allelic variation between individual humans occurs with a frequency of about once per each 500 bases. Much ofthe allelic variation is due to single nucleotide polymorphisms (SNPs), which include restriction fragment length polymorphisms (RFLPs).

Each of the sequences described herein can, to some degree, be used as a standard against which DNA from an individual can be compared for identification purposes. Because greater numbers of polymorphisms occur in the noncoding regions, fewer sequences are necessary to differentiate individuals. The noncoding sequences can comfortably provide positive individual identification with a panel of perhaps 10 to 1,000 primers that each yield a noncoding amplified sequence of 100 bases. If predicted coding sequences, such as those in SEQ LD NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47 are used, a more appropriate number of primers for positive individual identification would be 500-2,000.

Predictive Medicine

The invention also pertains to the field of predictive medicine in which diagnostic assays, prognostic assays, pharmacogenomics, and monitoring clinical trials are used for prognostic (predictive) purposes to thereby treat an individual prophylactically. Accordingly, one aspect ofthe invention relates to diagnostic assays for determining NOVX protein and/or nucleic acid expression as well as NOVX activity, in the context of a biological sample (e.g., blood, serum, cells, tissue) to thereby determine whether an individual is afflicted with a disease or disorder, or is at risk of developing a disorder, associated with aberrant NOVX expression or activity. The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers. The invention also provides for prognostic (or predictive) assays for determining whether an individual is at risk of developing a disorder associated with NOVX protein, nucleic acid expression or activity. For example, mutations in an NOVX gene can be assayed in a biological sample. Such assays can be used for prognostic or predictive purpose to thereby prophylactically treat an individual prior to the onset of a disorder characterized by or associated with NOVX protein, nucleic acid expression, or biological activity.

Another aspect ofthe invention provides methods for determining NOVX protein, nucleic acid expression or activity in an individual to thereby select appropriate therapeutic or prophylactic agents for that individual (referred to herein as "pharmacogenomics"). Pharmacogenomics allows for the selection of agents (e.g., dmgs) for therapeutic or prophylactic treatment of an individual based on the genotype ofthe individual (e.g., the genotype ofthe individual examined to determine the ability ofthe individual to respond to a particular agent.) Yet another aspect ofthe invention pertains to monitoring the influence of agents (e.g., d gs, compounds) on the expression or activity of NOVX in clinical trials.

These and other agents are described in further detail in the following sections.

Diagnostic Assays

An exemplary method for detecting the presence or absence of NOVX in a biological sample involves obtaining a biological sample from a test subject and contacting the biological sample with a compound or an agent capable of detecting NOVX protein or nucleic acid (e.g., mRNA, genomic DNA) that encodes NOVX protein such that the presence of NOVX is detected in the biological sample. An agent for detecting NOVX mRNA or genomic DNA is a labeled nucleic acid probe capable of hybridizing to NOVX mRNA or genomic DNA. The nucleic acid probe can be, for example, a full-length NOVX nucleic acid, such as the nucleic acid of SEQ LD NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and 47, or a portion thereof, such as an oligonucleotide of at least 15, 30, 50, 100, 250 or 500 nucleotides in length and sufficient to specifically hybridize under stringent conditions to NOVX mRNA or genomic DNA. Other suitable probes for use in the diagnostic assays of the invention are described herein.

An agent for detecting NOVX protein is an antibody capable of binding to NOVX protein, preferably an antibody with a detectable label. Antibodies can be polyclonal, or more preferably, monoclonal. An intact antibody, or a fragment thereof (e.g., Fab or F(ab')₂) can be used. The term "labeled", with regard to the probe or antibody, is intended to encompass direct labeling ofthe probe or antibody by coupling (t.e., physically linking) a detectable substance to the probe or antibody, as well as indirect labeling ofthe probe or antibody by reactivity with another reagent that is directly labeled. Examples of indirect labeling include detection of a primary antibody using a fluorescently- labeled secondary antibody and end-labeling of a DNA probe with biotin such that it can be detected with fluorescently- labeled streptavidin. The term "biological sample" is intended to include tissues, cells and biological fluids isolated from a subject, as well as tissues, cells and fluids present within a subject. That is, the detection method ofthe invention can be used to detect NOVX mRNA, protein, or genomic DNA in a biological sample in vitro as well as in vivo. For example, in vitro techniques for detection of NOVX mRNA include Northern hybridizations and in situ hybridizations. In vitro techniques for detection of NOVX protein include enzyme linked immunosorbent assays (ELISAs), Western blots, immunoprecipitations, and immunofluorescence. In vitro techniques for detection of NOVX genomic DNA include Southern hybridizations. Furthermore, in vivo techniques for detection of NOVX protein include introducing into a subject a labeled anti-NOVX antibody. For example, the antibody can be labeled with a radioactive marker whose presence and location in a subject can be detected by standard imaging techniques.

In one embodiment, the biological sample contains protein molecules from the test subject. Alternatively, the biological sample can contain mRNA molecules from the test subject or genomic DNA molecules from the test subject. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject.

In another embodiment, the methods further involve obtaining a control biological sample from a control subject, contacting the control sample with a compound or agent capable of detecting NOVX protein, mRNA, or genomic DNA, such that the presence of

NOVX protein, mRNA or genomic DNA is detected in the biological sample, and comparing the presence of NOVX protein, mRNA or genomic DNA in the control sample with the presence of NOVX protein, mRNA or genomic DNA in the test sample. The invention also encompasses kits for detecting the presence of NOVX in a biological sample. For example, the kit can comprise: a labeled compound or agent capable of detecting NOVX protein or mRNA in a biological sample; means for determining the amount of NOVX in the sample; and means for comparing the amount of NOVX in the sample with a standard. The compound or agent can be packaged in a suitable container. The kit can further comprise instmctions for using the kit to detect NOVX protein or nucleic acid.

Prognostic Assays

The diagnostic methods described herein can furthermore be utilized to identify subjects having or at risk of developing a disease or disorder associated with aberrant NOVX expression or activity. For example, the assays described herein, such as the preceding diagnostic assays or the following assays, can be utilized to identify a subject having or at risk of developing a disorder associated with NOVX protein, nucleic acid expression or activity. Alternatively, the prognostic assays can be utilized to identify a subject having or at risk for developing a disease or disorder. Thus, the invention provides a method for identifying a disease or disorder associated with aberrant NOVX expression or activity in which a test sample is obtained from a subject and NOVX protein or nucleic acid (e.g., mRNA, genomic DNA) is detected, wherein the presence of NOVX protein or nucleic acid is diagnostic for a subject having or at risk of developing a disease or disorder associated with aberrant NOVX expression or activity. As used herein, a "test sample" refers to a biological sample obtained from a subject of interest. For example, a test sample can be a biological fluid (e.g., semm), cell sample, or tissue.

Furthermore, the prognostic assays described herein can be used to determine whether a subject can be administered an agent (e.g., an agonist, antagonist, peptidomimetic, protein, peptide, nucleic acid, small molecule, or other dmg candidate) to treat a disease or disorder associated with aberrant NOVX expression or activity. For example, such methods can be used to determine whether a subject can be effectively treated with an agent for a disorder. Thus, the invention provides methods for determining whether a subject can be effectively treated with an agent for a disorder associated with aberrant NOVX expression or activity in which a test sample is obtained and NOVX protein or nucleic acid is detected (e.g., wherein the presence of NOVX protein or nucleic acid is diagnostic for a subject that can be administered the agent to treat a disorder associated with aberrant NOVX expression or activity).

The methods ofthe invention can also be used to detect genetic lesions in an NOVX gene, thereby determining if a subject with the lesioned gene is at risk for a disorder characterized by aberrant cell proliferation and/or differentiation. In various embodiments, the methods include detecting, in a sample of cells from the subject, the presence or absence of a genetic lesion characterized by at least one of an alteration affecting the integrity of a gene encoding an NOVX-protein, or the misexpression ofthe NOVX gene. For example, such genetic lesions can be detected by ascertaining the existence of at least one of: (i) a deletion of one or more nucleotides from an NOVX gene; (ii) an addition of one or more nucleotides to an NOVX gene; (ttt) a substitution of one or more nucleotides of an NOVX gene, (iv) a chromosomal rearrangement of an NOVX gene; (v) an alteration in the level of a messenger RNA transcript of an NOVX gene, (vt) aberrant modification of an NOVX gene, such as ofthe methylation pattern ofthe genomic DNA, (vtt) the presence of a non- wild-type splicing pattern of a messenger RNA transcript of an NOVX gene, (viii) a non- wild-type level of an NOVX protein, (ix) allelic loss of an NOVX gene, and (x) inappropriate post-translational modification of an NOVX protein. As described herein, there are a large number of assay techniques known in the art which can be used for detecting lesions in an NOVX gene. A preferred biological sample is a peripheral blood leukocyte sample isolated by conventional means from a subject. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.

In certain embodiments, detection ofthe lesion involves the use of a probe/primer in a polymerase chain reaction (PCR) (see, e.g., U.S. Patent Nos. 4,683,195 and 4,683,202), such as anchor PCR or RACE PCR, or, alternatively, in a ligation chain reaction (LCR) (see, e.g., Landegran, et αl., 1988. Science 241 : 1077-1080; and Nakazawa, et αl., 1994. Proc. Nαtl Acαd. Sci. USA 91: 360-364), the latter of which can be particularly useful for detecting point mutations in the NOVX-gene (see, Abravaya, et αl, 1995. Nucl. Acids Res. 23: 675-682). This method can include the steps of collecting a sample of cells from a patient, isolating nucleic acid (e.g., genomic, mRNA or both) from the cells ofthe sample, contacting the nucleic acid sample with one or more primers that specifically hybridize to an NOVX gene under conditions such that hybridization and amplification ofthe NOVX gene (if present) occurs, and detecting the presence or absence of an amplification product, or detecting the size ofthe amplification product and comparing the length to a control sample. It is anticipated that PCR and/or LCR may be desirable to use as a preliminary amplification step in conjunction with any ofthe techniques used for detecting mutations described herein.

Alternative amplification methods include: self sustained sequence replication (see, Guatelli, et αl, 1990. Proc. Nαtl. Acαd. Sci. USA 87: 1874-1878), transcriptional amplification system (see, Kwoh, et αl., 1989. Proc. Nαtl. Acαd. Sci. USA 86: 1173-1177); Qβ Replicase (see, Lizardi, et al, 1988. BioTechnology 6: 1197), or any other nucleic acid amplification method, followed by the detection ofthe amplified molecules using techniques well known to those of skill in the art. These detection schemes are especially useful for the detection of nucleic acid molecules if such molecules are present in very low numbers. In an alternative embodiment, mutations in an NOVX gene from a sample cell can be identified by alterations in restriction enzyme cleavage patterns. For example, sample and control DNA is isolated, amplified (optionally), digested with one or more restriction endonucleases, and fragment length sizes are determined by gel electrophoresis and compared. Differences in fragment length sizes between sample and control DNA indicates mutations in the sample DNA. Moreover, the use of sequence specific ribozymes (see, e.g., U.S. Patent No. 5,493,531) can be used to score for the presence of specific mutations by development or loss of a ribozyme cleavage site.

In other embodiments, genetic mutations in NOVX can be identified by hybridizing a sample and control nucleic acids, e.g., DNA or RNA, to high-density arrays containing hundreds or thousands of oligonucleotides probes. See, e.g., Cronin, et al, 1996. Human Mutation 7: 244-255; Kozal, et al, 1996. Nat. Med. 2: 753-759. For example, genetic mutations in NOVX can be identified in two dimensional arrays containing light-generated DNA probes as described in Cronin, et al, supra. Briefly, a first hybridization array of probes can be used to scan through long stretches of DNA in a sample and control to identify base changes between the sequences by making linear arrays of sequential overlapping probes. This step allows the identification of point mutations. This is followed by a second hybridization array that allows the characterization of specific mutations by using smaller, specialized probe arrays complementary to all variants or mutations detected. Each mutation array is composed of parallel probe sets, one complementary to the wild- type gene and the other complementary to the mutant gene.

In yet another embodiment, any of a variety of sequencing reactions known in the art can be used to directly sequence the NOVX gene and detect mutations by comparing the sequence ofthe sample NOVX with the corresponding wild-type (control) sequence. Examples of sequencing reactions include those based on techniques developed by Maxim and Gilbert, 1977. Proc. Natl. Acad. Sci. USA 74: 560 or Sanger, 1977. Proc. Natl. Acad. Sci. USA 74: 5463. It is also contemplated that any of a variety of automated sequencing procedures can be utilized when performing the diagnostic assays (see, e.g., Naeve, et al, 1995. Biotechniques 19: 448), including sequencing by mass spectrometry (see, e.g., PCT International Publication No. WO 94/16101; Cohen, et al, 1996. Adv. Chromatography 36: 127-162; and Griffin, et al, 1993. Appl. Biochem. Biotechnol. 38: 147-159).

Other methods for detecting mutations in the NOVX gene include methods in which protection from cleavage agents is used to detect mismatched bases in RNA/RNA or RNA/DNA heteroduplexes. See, e.g., Myers, et al, 1985. Science 230: 1242. In general, the art technique of "mismatch cleavage" starts by providing heteroduplexes of formed by hybridizing (labeled) RNA or DNA containing the wild-type NOVX sequence with potentially mutant RNA or DNA obtained from a tissue sample. The double-stranded duplexes are treated with an agent that cleaves single-stranded regions ofthe duplex such as which will exist due to basepair mismatches between the control and sample strands. For instance, RNA/DNA duplexes can be treated with RNase and DNA/DNA hybrids treated with Sj nuclease to enzymatically digesting the mismatched regions. In other embodiments, either DNA/DNA or RNA/DNA duplexes can be treated with hydroxylamine or osmium tetroxide and with piperidine in order to digest mismatched regions. After digestion ofthe mismatched regions, the resulting material is then separated by size on denaturing polyacrylamide gels to determine the site of mutation. See, e.g., Cotton, et al, 1988. Proc. Natl. Acad. Sci. USA 85: 4397; Saleeba, et al, 1992. Methods Enzymol. 217: 286-295. In an embodiment, the control DNA or RNA can be labeled for detection.

In still another embodiment, the mismatch cleavage reaction employs one or more proteins that recognize mismatched base pairs in double-stranded DNA (so called "DNA mismatch repair" enzymes) in defined systems for detecting and mapping point mutations in NOVX cDNAs obtained from samples of cells. For example, the mutY enzyme of E. coli cleaves A at G/A mismatches and the thymidine DNA glycosylase from HeLa cells cleaves T at G/T mismatches. See, e.g., Hsu, et al, 1994. Carcinogenesis 15: 1657-1662. According to an exemplary embodiment, a probe based on an NOVX sequence, e.g., a wild-type NOVX sequence, is hybridized to a cDNA or other DNA product from a test cell(s). The duplex is treated with a DNA mismatch repair enzyme, and the cleavage products, if any, can be detected from electrophoresis protocols or the like. See, e.g., U.S. Patent No. 5,459,039.

In other embodiments, alterations in electrophoretic mobility will be used to identify mutations in NOVX genes. For example, single strand conformation polymorphism (SSCP) may be used to detect differences in electrophoretic mobility between mutant and wild type nucleic acids. See, e.g., Orita, et al, 1989. Proc. Natl. Acad. Sci. USA: 86: 2766; Cotton, 1993. Mutat. Res. 285: 125-144; Hayashi, 1992. Genet. Anal. Tech. Appl. 9: 73-79. Single-stranded DNA fragments of sample and control NOVX nucleic acids will be denatured and allowed to renature. The secondary stmcture of single-stranded nucleic acids varies according to sequence, the resulting alteration in electrophoretic mobility enables the detection of even a single base change. The DNA fragments may be labeled or detected with labeled probes. The sensitivity ofthe assay may be enhanced by using RNA (rather than DNA), in which the secondary stmcture is more sensitive to a change in sequence. In one embodiment, the subject method utilizes heteroduplex analysis to separate double stranded heteroduplex molecules on the basis of changes in electrophoretic mobility. See, e.g., Keen, et al, 1991. Trends Genet. 1: 5.

In yet another embodiment, the movement of mutant or wild-type fragments in polyacrylamide gels containing a gradient of denaturant is assayed using denaturing gradient gel electrophoresis (DGGE). See, e.g., Myers, et al, 1985. Nature 313: 495. When DGGE is used as the method of analysis, DNA will be modified to insure that it does not completely denature, for example by adding a GC clamp of approximately 40 bp of high-melting GC-rich DNA by PCR. In a further embodiment, a temperature gradient is used in place of a denaturing gradient to identify differences in the mobility of control and sample DNA. See, e.g., Rosenbaum and Reissner, 1987. Biophys. Chem. 265: 12753.

Examples of other techniques for detecting point mutations include, but are not limited to, selective oligonucleotide hybridization, selective amplification, or selective primer extension. For example, oligonucleotide primers may be prepared in which the known mutation is placed centrally and then hybridized to target DNA under conditions that permit hybridization only if a perfect match is found. See, e.g., Saiki, et al, 1986. Nature 324: 163; Saiki, et al, 1989. Proc. Natl Acad. Sci. USA 86: 6230. Such allele specific oligonucleotides are hybridized to PCR amplified target DNA or a number of different mutations when the oligonucleotides are attached to the hybridizing membrane and hybridized with labeled target DNA.

Alternatively, allele specific amplification technology that depends on selective PCR amplification may be used in conjunction with the instant invention. Oligonucleotides used as primers for specific amplification may carry the mutation of interest in the center ofthe molecule (so that amplification depends on differential hybridization; see, e.g., Gibbs, et al, 1989. Nucl. Acids Res. 17: 2437-2448) or at the extreme 3'-terminus of one primer where, under appropriate conditions, mismatch can prevent, or reduce polymerase extension (see, e.g., Prossner, 1993. Tibtech. 11 : 238). In addition it may be desirable to introduce a novel restriction site in the region ofthe mutation to create cleavage-based detection. See, e.g., Gasparini, et al, 1992. Mol. Cell Probes 6: 1. It is anticipated that in certain embodiments amplification may also be performed using Taq ligase for amplification. See, e.g., Barany, 1991. Proc. Natl. Acad. Sci. USA 88: 189. In such cases, ligation will occur only if there is a perfect match at the 3'-terminus ofthe 5' sequence, making it possible to detect the presence of a known mutation at a specific site by looking for the presence or absence of amplification. The methods described herein may be performed, for example, by utilizing pre-packaged diagnostic kits comprising at least one probe nucleic acid or antibody reagent described herein, which may be conveniently used, e.g., in clinical settings to diagnose patients exhibiting symptoms or family history of a disease or illness involving an NOVX gene. Furthermore, any cell type or tissue, preferably peripheral blood leukocytes, in which

NOVX is expressed may be utilized in the prognostic assays described herein. However, any biological sample containing nucleated cells may be used, including, for example, buccal mucosal cells.

Pharmacogenomics Agents, or modulators that have a stimulatory or inhibitory effect on NOVX activity

(e.g., NOVX gene expression), as identified by a screening assay described herein can be administered to individuals to treat (prophylactically or therapeutically) disorders (The disorders include metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer- associated cachexia, cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, and hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.) In conjunction with such treatment, the pharmacogenomics (i.e., the study ofthe relationship between an individual's genotype and that individual's response to a foreign compound or drug) ofthe individual may be considered. Differences in metabolism of therapeutics can lead to severe toxicity or therapeutic failure by altering the relation between dose and blood concentration ofthe pharmacologically active dmg. Thus, the pharmacogenomics ofthe individual permits the selection of effective agents (e.g., dmgs) for prophylactic or therapeutic treatments based on a consideration ofthe individual's genotype. Such pharmacogenomics can further be used to determine appropriate dosages and therapeutic regimens. Accordingly, the activity of NOVX protein, expression of NOVX nucleic acid, or mutation content of NOVX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment ofthe individual. Pharmacogenomics deals with clinically significant hereditary variations in the response to dmgs due to altered dmg disposition and abnormal action in affected persons. See e.g., Eichelbaum, 1996. Clin. Exp. Pharmacol. Physiol, 23: 983-985; Linder, 1997. Clin. Chem., 43: 254-266. In general, two types of pharmacogenetic conditions can be differentiated. Genetic conditions transmitted as a single factor altering the way drugs act on the body (altered dmg action) or genetic conditions transmitted as single factors altering the way the body acts on drugs (altered dmg metabolism). These pharmacogenetic conditions can occur either as rare defects or as polymorphisms. For example, glucose-6-phosphate dehydrogenase (G6PD) deficiency is a common inherited enzymopathy in which the main clinical complication is hemolysis after ingestion of oxidant dmgs (anti-malarials, sulfonamides, analgesics, nitrofurans) and consumption of fava beans.

As an illustrative embodiment, the activity of dmg metabolizing enzymes is a major determinant of both the intensity and duration of dmg action. The discovery of genetic polymorphisms of dmg metabolizing enzymes (e.g., N-acetyltransferase 2 (NAT 2) and cytochrome P450 enzymes CYP2D6 and CYP2C19) has provided an explanation as to why some patients do not obtain the expected dmg effects or show exaggerated dmg response and serious toxicity after taking the standard and safe dose of a dmg. These polymorphisms are expressed in two phenotypes in the population, the extensive metabolizer (EM) and poor metabolizer (PM). The prevalence of PM is different among different populations. For example, the gene coding for CYP2D6 is highly polymorphic and several mutations have been identified in PM, which all lead to the absence of functional CYP2D6. Poor metabolizers of CYP2D6 and CYP2C19 quite frequently experience exaggerated dmg response and side effects when they receive standard doses. If a metabolite is the active therapeutic moiety, PM show no therapeutic response, as demonstrated for the analgesic effect of codeine mediated by its CYP2D6-formed metabolite morphine. At the other extreme are the so called ultra-rapid metabolizers who do not respond to standard doses. Recently, the molecular basis of ultra-rapid metabolism has been identified to be due to CYP2D6 gene amplification.

Thus, the activity of NOVX protein, expression of NOVX nucleic acid, or mutation content of NOVX genes in an individual can be determined to thereby select appropriate agent(s) for therapeutic or prophylactic treatment ofthe individual. In addition, pharmacogenetic studies can be used to apply genotyping of polymorphic alleles encoding drug-metabolizing enzymes to the identification of an individual's drug responsiveness phenotype. This knowledge, when applied to dosing or dmg selection, can avoid adverse reactions or therapeutic failure and thus enhance therapeutic or prophylactic efficiency when treating a subject with an NOVX modulator, such as a modulator identified by one ofthe exemplary screening assays described herein.

Monitoring of Effects During Clinical Trials

Monitoring the influence of agents (e.g., dmgs, compounds) on the expression or activity of NOVX (e.g. , the ability to modulate aberrant cell. proliferation and/or differentiation) can be applied not only in basic dmg screening, but also in clinical trials. For example, the effectiveness of an agent determined by a screening assay as described herein to increase NOVX gene expression, protein levels, or upregulate NOVX activity, can be monitored in clinical trails of subjects exhibiting decreased NOVX gene expression, protein levels, or downregulated NOVX activity. Alternatively, the effectiveness of an agent determined by a screening assay to decrease NOVX gene expression, protein levels, or downregulate NOVX activity, can be monitored in clinical trails of subjects exhibiting increased NOVX gene expression, protein levels, or upregulated NOVX activity. In such clinical trials, the expression or activity of NOVX and, preferably, other genes that have been implicated in, for example, a cellular proliferation or immune disorder can be used as a "read out" or markers ofthe immune responsiveness of a particular cell.

By way of example, and not of limitation, genes, including NOVX, that are modulated in cells by treatment with an agent (e.g., compound, dmg or small molecule) that modulates NOVX activity (e.g., identified in a screening assay as described herein) can be identified. Thus, to study the effect of agents on cellular proliferation disorders, for example, in a clinical trial, cells can be isolated and RNA prepared and analyzed for the levels of expression of NOVX and other genes implicated in the disorder. The levels of gene expression (i.e., a gene expression pattern) can be quantified by Northern blot analysis or RT-PCR, as described herein, or alternatively by measuring the amount of protein produced, by one ofthe methods as described herein, or by measuring the levels of activity of NOVX or other genes. In this manner, the gene expression pattern can serve as a marker, indicative ofthe physiological response ofthe cells to the agent. Accordingly, this response state may be determined before, and at various points during, treatment ofthe individual with the agent.

In one embodiment, the invention provides a method for monitoring the effectiveness of treatment of a subject with an agent (e.g., an agonist, antagonist, protein, peptide, peptidomimetic, nucleic acid, small molecule, or other dmg candidate identified by the screening assays described herein) comprising the steps of ( ) obtaining a pre-administration sample from a subject prior to administration ofthe agent; (ii) detecting the level of expression of an NOVX protein, mRNA, or genomic DNA in the preadminisfration sample; (ttt) obtaining one or more post-administration samples from the subject; (tv) detecting the level of expression or activity ofthe NOVX protein, mRNA, or genomic DNA in the post-administration samples; (v) comparing the level of expression or activity ofthe NOVX protein, mRNA, or genomic DNA in the pre-administration sample with the NOVX protein, mRNA, or genomic DNA in the post administration sample or samples; and (vt) altering the administration ofthe agent to the subject accordingly. For example, increased administration ofthe agent may be desirable to increase the expression or activity of NOVX to higher levels than detected, i.e., to increase the effectiveness ofthe agent. Alternatively, decreased administration ofthe agent may be desirable to decrease expression or activity of NOVX to lower levels than detected, t.e., to decrease the effectiveness ofthe agent.

Methods of Treatment

The invention provides for both prophylactic and therapeutic methods of treating a subject at risk of (or susceptible to) a disorder or having a disorder associated with aberrant NOVX expression or activity. The disorders include cardiomyopathy, atherosclerosis, hypertension, congenital heart defects, aortic stenosis, atrial septal defect (ASD), atrioventricular (A-V) canal defect, ductus arteriosus, pulmonary stenosis, subaortic stenosis, ventricular septal defect (VSD), valve diseases, tuberous sclerosis, scleroderma, obesity, transplantation, adrenoleukodystrophy, congenital adrenal hypeφlasia, prostate cancer, neoplasm; adenocarcinoma, lymphoma, utems cancer, fertility, hemophilia, hypercoagulation, idiopathic thrombocytopenic puφura, immunodeficiencies, graft versus host disease, ALDS, bronchial asthma, Crohn's disease; multiple sclerosis, treatment of Albright Hereditary Ostoeodystrophy, and other diseases, disorders and conditions ofthe like. These methods of treatment will be discussed more fully, below.

Disease and Disorders Diseases and disorders that are characterized by increased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that antagonize (i.e., reduce or inhibit) activity. Therapeutics that antagonize activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to: (i) an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof; (ii) antibodies to an aforementioned peptide; (ttt) nucleic acids encoding an aforementioned peptide; (tv) administration of antisense nucleic acid and nucleic acids that are "dysfunctional" (t.e., due to a heterologous insertion within the coding sequences of coding sequences to an aforementioned peptide) that are utilized to "knockout" endogenous function of an aforementioned peptide by homologous recombination (see, e.g., Capecchi, 1989. Science 244: 1288-1292); or (v) modulators ( t.e., inhibitors, agonists and antagonists, including additional peptide mimetic ofthe invention or antibodies specific to a peptide ofthe invention) that alter the interaction between an aforementioned peptide and its binding partner.

Diseases and disorders that are characterized by decreased (relative to a subject not suffering from the disease or disorder) levels or biological activity may be treated with Therapeutics that increase (i.e., are agonists to) activity. Therapeutics that upregulate activity may be administered in a therapeutic or prophylactic manner. Therapeutics that may be utilized include, but are not limited to, an aforementioned peptide, or analogs, derivatives, fragments or homologs thereof; or an agonist that increases bioavailability.

Increased or decreased levels can be readily detected by quantifying peptide and/or RNA, by obtaining a patient tissue sample (e.g., from biopsy tissue) and assaying it in vitro for RNA or peptide levels, stmcture and/or activity ofthe expressed peptides (or mRNAs of an aforementioned peptide). Methods that are well-known within the art include, but are not limited to, immunoassays (e.g., by Western blot analysis, immunoprecipitation followed by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis, immunocytochemistry, etc.) and/or hybridization assays to detect expression of mRNAs (e.g., Northern assays, dot blots, in situ hybridization, and the like). Prophylactic Methods

In one aspect, the invention provides a method for preventing, in a subject, a disease or condition associated with an aberrant NOVX expression or activity, by administering to the subject an agent that modulates NOVX expression or at least one NOVX activity. Subjects at risk for a disease that is caused or contributed to by aberrant NOVX expression or activity can be identified by, for example, any or a combination of diagnostic or prognostic assays as described herein. Administration of a prophylactic agent can occur prior to the manifestation of symptoms characteristic ofthe NOVX aberrancy, such that a disease or disorder is prevented or, alternatively, delayed in its progression. Depending upon the type of NOVX aberrancy, for example, an NOVX agonist or NOVX antagonist agent can be used for treating the subject. The appropriate agent can be determined based on screening assays described herein. The prophylactic methods ofthe invention are further discussed in the following subsections.

Therapeutic Methods Another aspect ofthe invention pertains to methods of modulating NOVX expression or activity for therapeutic p poses. The modulatory method ofthe invention involves contacting a cell with an agent that modulates one or more ofthe activities of NOVX protein activity associated with the cell. An agent that modulates NOVX protein activity can be an agent as described herein, such as a nucleic acid or a protein, a naturally-occurring cognate ligand of an NOVX protein, a peptide, an NOVX peptidomimetic, or other small molecule. In one embodiment, the agent stimulates one or more NOVX protein activity. Examples of such stimulatory agents include active NOVX protein and a nucleic acid molecule encoding NOVX that has been introduced into the cell. In another embodiment, the agent inhibits one or more NOVX protein activity. Examples of such inhibitory agents include antisense NOVX nucleic acid molecules and anti-NOVX antibodies. These modulatory methods can be performed in vitro (e.g., by culturing the cell with the agent) or, alternatively, in vivo (e.g., by administering the agent to a subject). As such, the invention provides methods of treating an individual afflicted with a disease or disorder characterized by aberrant expression or activity of an NOVX protein or nucleic acid molecule. In one embodiment, the method involves administering an agent (e.g., an agent identified by a screening assay described herein), or combination of agents that modulates (e.g., up-regulates or down-regulates) NOVX expression or activity. In another embodiment, the method involves administering an NOVX protein or nucleic acid molecule as therapy to compensate for reduced or aberrant NOVX expression or activity.

Stimulation of NOVX activity is desirable in sttwations in which NOVX is abnormally downregulated and/or in which increased NOVX activity is likely to have a beneficial effect. One example of such a situation is where a subject has a disorder characterized by aberrant cell proliferation and/or differentiation (e.g., cancer or immune associated disorders). Another example of such a situation is where the subject has a gestational disease (e.g., preclampsia).

Determination ofthe Biological Effect of the Therapeutic

In various embodiments ofthe invention, suitable in vitro or in vivo assays are performed to determine the effect of a specific Therapeutic and whether its administration is indicated for treatment ofthe affected tissue. In various specific embodiments, in vitro assays may be performed with representative cells ofthe type(s) involved in the patient's disorder, to determine if a given Therapeutic exerts the desired effect upon the cell type(s). Compounds for use in therapy may be tested in suitable animal model systems including, but not limited to rats, mice, chicken, cows, monkeys, rabbits, and the like, prior to testing in human subjects. Similarly, for in vivo testing, any ofthe animal model system known in the art may be used prior to administration to human subjects.

Prophylactic and Therapeutic Uses of the Compositions of the Invention

The NOVX nucleic acids and proteins ofthe invention are useful in potential prophylactic and therapeutic applications implicated in a variety of disorders including, but not limited to: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer- associated cancer, neurodegenerative disorders, Alzheimer's Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyslipidemias, metabolic disturbances associated with obesity, the metabolic syndrome X and wasting disorders associated with chronic diseases and various cancers.

As an example, a cDNA encoding the NOVX protein ofthe invention may be useful in gene therapy, and the protein may be useful when administered to a subject in need thereof. By way of non-limiting example, the compositions ofthe invention will have efficacy for treatment of patients suffering from: metabolic disorders, diabetes, obesity, infectious disease, anorexia, cancer-associated cachexia, cancer, neurodegenerative disorders, Alzheimer's

Disease, Parkinson's Disorder, immune disorders, hematopoietic disorders, and the various dyslipidemias.

Both the novel nucleic acid encoding the NOVX protein, and the NOVX protein ofthe invention, or fragments thereof, may also be useful in diagnostic applications, wherein the presence or amount ofthe nucleic acid or the protein are to be assessed. A further use could be as an anti-bacterial molecule (t.e., some peptides have been found to possess anti-bacterial properties). These materials are further useful in the generation of antibodies, which immunospecifically-bind to the novel substances ofthe invention for use in therapeutic or diagnostic methods. The invention will be further described in the following examples, which do not limit the scope ofthe invention described in the claims.

Examples

EXAMPLE 1: Identification of NOVX Nucleic Acids

TblastN using CuraGen Coφoration's sequence file for polypeptides or homologs was n against the Genomic Daily Files made available by GenBank or from files downloaded from the individual sequencing centers. Exons were predicted by homology and the intron/exon boundaries were determined using standard genetic mles. Exons were further selected and refined by means of similarity determination using multiple BLAST (for example, tBlastN, BlastX, and BlastN) searches, and, in some instances, GeneScan and Grail. Expressed sequences from both public and proprietary databases were also added when available to further define and complete the gene sequence. The DNA sequence was then manually corrected for apparent inconsistencies thereby obtaining the sequences encoding the full-length protein.

The novel NOVX target sequences identified in the present invention were subjected to the exon linking process to confirm the sequence. PCR primers were designed by starting at the most upstream sequence available, for the forward primer, and at the most downstream sequence available for the reverse primer. PCR primer sequences were used for obtaining different clones. In each case, the sequence was examined, walking inward from the respective termini toward the coding sequence, until a suitable sequence that is either unique or highly selective was encountered, or, in the case ofthe reverse primer, until the stop codon was reached. Such primers were designed based on in silico predictions for the full length cDNA, part (one or more exons) ofthe DNA or protein sequence ofthe target sequence, or by translated homology ofthe predicted exons to closely related human sequences from other species. These primers were then employed in PCR amplification based on the following pool of human cDNAs: adrenal gland, bone marrow, brain - amygdala, brain - cerebellum, brain - hippocampus, brain - substantia nigra, brain - thalamus, brain -whole, fetal brain, fetal kidney, fetal liver, fetal lung, heart, kidney, lymphoma - Raji, mammary gland, pancreas, pituitary gland, placenta, prostate, salivary gland, skeletal muscle, small intestine, spinal cord, spleen, stomach, testis, thyroid, trachea, utems. Usually the resulting amplicons were gel purified, cloned and sequenced to high redundancy. The PCR product derived from exon linking was cloned into the pCR2.1 vector from Invitrogen. The resulting bacterial clone has an insert covering the entire open reading frame cloned into the pCR2.1 vector. The resulting sequences from all clones were assembled with themselves, with other fragments in CuraGen

Coφoration's database and with public ESTs. Fragments and ESTs were included as components for an assembly when the extent of their identity with another component ofthe assembly was at least 95% over 50 bp. In addition, sequence traces were evaluated manually and edited for corrections if appropriate. These procedures provide the sequence reported herein.

Physical clone: Exons were predicted by homology and the intron/exon boundaries were determined using standard genetic mles. Exons were further selected and refined by means of similarity determination using multiple BLAST (for example, tBlastN, BlastX, and BlastN) searches, and, in some instances, GeneScan and Grail. Expressed sequences from both public and proprietary databases were also added when available to further define and complete the gene sequence. The DNA sequence was then manually corrected for apparent inconsistencies thereby obtaining the sequences encoding the full-length protein.

Example 2: Identification of Single Nucleotide Polymorphisms in NOVX nucleic acid sequences

Variant sequences are also included in this application. A variant sequence can include a single nucleotide polymoφhism (SNP). A SNP can, in some instances, be referred to as a "cSNP" to denote that the nucleotide sequence containing the SNP originates as a cDNA. A SNP can arise in several ways. For example, a SNP may be due to a substitution of one nucleotide for another at the polymoφhic site. Such a substitution can be either a transition or a transversion. A SNP can also arise from a deletion of a nucleotide or an insertion of a nucleotide, relative to a reference allele. In this case, the polymoφhic site is a site at which one allele bears a gap with respect to a particular nucleotide in another allele. SNPs occurring within genes may result in an alteration ofthe amino acid encoded by the gene at the position ofthe SNP. Intragenic SNPs may also be silent, when a codon including a SNP encodes the same amino acid as a result ofthe redundancy ofthe genetic code. SNPs occurring outside the region of a gene, or in an intron within a gene, do not result in changes in any amino acid sequence of a protein but may result in altered regulation ofthe expression pattern. Examples include alteration in temporal expression, physiological response regulation, cell type expression regulation, intensity of expression, and stability of transcribed message.

SeqCalling assemblies produced by the exon linking process were selected and extended using the following criteria. Genomic clones having regions with 98% identity to all or part ofthe initial or extended sequence were identified by BLASTN searches using the relevant sequence to query human genomic databases. The genomic clones that resulted were selected for further analysis because this identity indicates that these clones contain the genomic locus for these SeqCalling assemblies. These sequences were analyzed for putative coding regions as well as for similarity to the known DNA and protein sequences. Programs used for these analyses include Grail, Genscan, BLAST, HMMER, FASTA, Hybrid and other relevant programs. Some additional genomic regions may have also been identified because selected

SeqCalling assemblies map to those regions. Such SeqCalling sequences may have overlapped with regions defined by homology or exon prediction. They may also be included because the location ofthe fragment was in the vicinity of genomic regions identified by similarity or exon prediction that had been included in the original predicted sequence. The sequence so identified was manually assembled and then may have been extended using one or more additional sequences taken from CuraGen Coφoration's human SeqCalling database. SeqCalling fragments suitable for inclusion were identified by the CuraTools™ program SeqExtend or by identifying SeqCalling fragments mapping to the appropriate regions ofthe genomic clones analyzed.

The regions defined by the procedures described above were then manually integrated and corrected for apparent inconsistencies that may have arisen, for example, from miscalled bases in the original fragments or from discrepancies between predicted exon junctions, EST locations and regions of sequence similarity, to derive the final sequence disclosed herein. When necessary, the process to identify and analyze SeqCalling assemblies and genomic clones was reiterated to derive the full length sequence.

Example 3. Quantitative expression analysis of clones in various cells and tissues

The quantitative expression of various clones was assessed using microtiter plates containing RNA samples from a variety of normal and pathology-derived cells, cell lines and tissues using real time quantitative PCR (RTQ PCR). RTQ PCR was performed on an

Applied Biosystems ABI PRISM® 7700 or an ABI PRISM® 7900 HT Sequence Detection System. Various collections of samples are assembled on the plates, and referred to as Panel 1 (containing normal tissues and cancer cell lines), Panel 2 (containing samples derived from tissues from normal and cancer sources), Panel 3 (containing cancer cell lines), Panel 4 (containing cells and cell lines from normal tissues and cells related to inflammatory conditions), Panel 5D/5I (containing human tissues and cell lines with an emphasis on metabolic diseases), AI_comprehensive_panel (containing normal tissue and samples from autoinflammatory diseases), Panel CNSD.01 (containing samples from normal and diseased brains) and CNS_neurodegeneration_panel (containing samples from normal and Alzheimer's diseased brains).

RNA integrity from all samples is controlled for quality by visual assessment of agarose gel electropherograms using 28S and 18S ribosomal RNA staining intensity ratio as a guide (2:1 to 2.5:1 28s:18s) and the absence of low molecular weight RNAs that would be indicative of degradation products. Samples are controlled against genomic DNA contamination by RTQ PCR reactions run in the absence of reverse transcriptase using probe and primer sets designed to amplify across the span of a single exon.

First, the RNA samples were normalized to reference nucleic acids such as constitutively expressed genes (for example, β-actin and GAPDH). Normalized RNA (5 ul) was converted to cDNA and analyzed by RTQ-PCR using One Step RT-PCR Master Mix Reagents (Applied Biosystems; Catalog No. 4309169) and gene-specific primers according to the manufacturer's instructions.

In other cases, non-normalized RNA samples were converted to single strand cDNA (sscDNA) using Superscript II (Invitrogen Coφoration; Catalog No. 18064-147) and random hexamers according to the manufacturer's instmctions. Reactions containing up to 10 μg of total RNA were performed in a volume of 20 μl and incubated for 60 minutes at 42°C. This reaction can be scaled up to 50 μg of total RNA in a final volume of 100 μl. sscDNA samples are then normalized to reference nucleic acids as described previously, using IX TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instmctions.

Probes and primers were designed for each assay according to Applied Biosystems Primer Express Software package (version I for Apple Computer's Macintosh Power PC) or a similar algorithm using the target sequence as input. Default settings were used for reaction conditions and the following parameters were set before selecting primers: primer concentration = 250 nM, primer melting temperature (T_m) range = 58°-60° C, primer optimal Tm = 59° C, maximum primer difference = 2° C, probe does not have 5' G, probe T_m must be 10° C greater than primer T_m, amplicon size 75 bp to 100 bp. The probes and primers selected (see below) were synthesized by Synthegen (Houston, TX, USA). Probes were double purified by HPLC to remove uncoupled dye and evaluated by mass spectroscopy to verify coupling of reporter and quencher dyes to the 5' and 3' ends ofthe probe, respectively. Their final concentrations were: forward and reverse primers, 900 nM each, and probe, 200nM. PCR conditions: When working with RNA samples, normalized RNA from each tissue and each cell line was spotted in each well of either a 96 well or a 384-well PCR plate (Applied Biosystems). PCR cocktails included either a single gene specific probe and primers set, or two multiplexed probe and primers sets (a set specific for the target clone and another gene-specific set multiplexed with the target probe). PCR reactions were set up using TaqMan® One-Step RT-PCR Master Mix (Applied Biosystems, Catalog No. 4313803) following manufacturer's instmctions. Reverse transcription was performed at 48° C for 30 minutes followed by amplification/PCR cycles as follows: 95° C 10 min, then 40 cycles of 95° C for 15 seconds, 60° C for 1 minute. Results were recorded as CT values (cycle at which a given sample crosses a threshold level of fluorescence) using a log scale, with the difference in RNA concentration between a given sample and the sample with the lowest CT value being represented as 2 to the power of delta CT. The percent relative expression is then obtained by taking the reciprocal of this RNA difference and multiplying by 100. When working with sscDNA samples, normalized sscDNA was used as described previously for RNA samples. PCR reactions containing one or two sets of probe and primers were set up as described previously, using IX TaqMan® Universal Master mix (Applied Biosystems; catalog No. 4324020), following the manufacturer's instmctions. PCR amplification was performed as follows: 95° C 10 min, then 40 cycles of 95° C for 15 seconds, 60° C for 1 minute. Results were analyzed and processed as described previously.

Panels 1, 1.1, 1.2, and 1.3D

The plates for Panels 1, 1.1, 1.2 and 1.3D include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in these panels are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers ofthe following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in these panels are widely available through the American Type Culture Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on these panels are comprised of samples derived from all major organ systems from single adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions ofthe brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, utems, placenta, prostate, testis and adipose.

In the results for Panels 1, 1.1, 1.2 and 1.3D, the following abbreviations are used: ca. = carcinoma, * = established from metastasis, met = metastasis, s cell var = small cell variant, non-s = non-sm = non-small, squam = squamous, pi. eff = pi effusion = pleural effusion, glio = glioma, astro = astrocytoma, and neuro = neuroblastoma.

General_screening_panel_vl.4 The plates for Panel 1.4 include 2 control wells (genomic DNA control and chemistry control) and 94 wells containing cDNA from various samples. The samples in Panel 1.4 are broken into 2 classes: samples derived from cultured cell lines and samples derived from primary normal tissues. The cell lines are derived from cancers ofthe following types: lung cancer, breast cancer, melanoma, colon cancer, prostate cancer, CNS cancer, squamous cell carcinoma, ovarian cancer, liver cancer, renal cancer, gastric cancer and pancreatic cancer. Cell lines used in Panel 1.4 are widely available through the American Type Culture

Collection (ATCC), a repository for cultured cell lines, and were cultured using the conditions recommended by the ATCC. The normal tissues found on Panel 1.4 are comprised of pools of samples derived from all major organ systems from 2 to 5 different adult individuals or fetuses. These samples are derived from the following organs: adult skeletal muscle, fetal skeletal muscle, adult heart, fetal heart, adult kidney, fetal kidney, adult liver, fetal liver, adult lung, fetal lung, various regions ofthe brain, the spleen, bone marrow, lymph node, pancreas, salivary gland, pituitary gland, adrenal gland, spinal cord, thymus, stomach, small intestine, colon, bladder, trachea, breast, ovary, ute s, placenta, prostate, testis and adipose. Abbreviations are as described for Panels 1, 1.1, 1.2, and 1.3D. Panels 2D and 2.2

The plates for Panels 2D and 2.2 generally include 2 control wells and 94 test samples composed of RNA or cDNA isolated from human tissue procured by surgeons working in close cooperation with the National Cancer Institute's Cooperative Human Tissue Network (CHTN) or the National Disease Research Initiative (NDRI). The tissues are derived from human malignancies and in cases where indicated many malignant tissues have "matched margins" obtained from noncancerous tissue just adjacent to the tumor. These are termed normal adjacent tissues and are denoted "NAT" in the results below. The tumor tissue and the "matched margins" are evaluated by two independent pathologists (the surgical pathologists and again by a pathologist at NDRI or CHTN). This analysis provides a gross histopatho logical assessment of tumor differentiation grade. Moreover, most samples include the original surgical pathology report that provides information regarding the clinical stage of the patient. These matched margins are taken from the tissue surrounding (i.e. immediately proximal) to the zone of surgery (designated "NAT", for normal adjacent tissue, in Table RR). In addition, RNA and cDNA samples were obtained from various human tissues derived from autopsies performed on elderly people or sudden death victims (accidents, etc.). These tissues were ascertained to be free of disease and were purchased from various commercial sources such as Clontech (Palo Alto, CA), Research Genetics, and Invitrogen. Panel 3D

The plates of Panel 3D are comprised of 94 cDNA samples and two control samples. Specifically, 92 of these samples are derived from cultured human cancer cell lines, 2 samples of human primary cerebellar tissue and 2 controls. The human cell lines are generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: Squamous cell carcinoma ofthe tongue, breast cancer, prostate cancer, melanoma, epidermoid carcinoma, sarcomas, bladder carcinomas, pancreatic cancers, kidney cancers, leukemias/lymphomas, ovarian/uterine/cervical, gastric, colon, lung and CNS cancer cell lines. In addition, there are two independent samples of cerebellum. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. The cell lines in panel 3D and 1.3D are ofthe most common cell lines used in the scientific literature.

Panels 4D, 4R, and 4.1D

Panel 4 includes samples on a 96 well plate (2 control wells, 94 test samples) composed of RNA (Panel 4R) or cDNA (Panels 4D/4.1 D) isolated from various human cell lines or tissues related to inflammatory conditions. Total RNA from control normal tissues such as colon and lung (Stratagene, La Jolla, CA) and thymus and kidney (Clontech) was employed. Total RNA from liver tissue from cirrhosis patients and kidney from lupus patients was obtained from BioChain (Biochain Institute, Inc., Hayward, CA). Intestinal tissue for RNA preparation from patients diagnosed as having Crohn's disease and ulcerative colitis was obtained from the National Disease Research Interchange (NDRI) (Philadelphia, PA).

Astrocytes, lung fibroblasts, dermal fibroblasts, coronary artery smooth muscle cells, small airway epithelium, bronchial epithelium, microvascular dermal endothelial cells, microvascular lung endothelial cells, human pulmonary aortic endothelial cells, human umbilical vein endothelial cells were all purchased from Clonetics (Walkersville, MD) and grown in the media supplied for these cell types by Clonetics. These primary cell types were activated with various cytokines or combinations of cytokines for 6 and/or 12-14 hours, as indicated. The following cytokines were used; IL-1 beta at approximately 1-5 ng/ml, TNF alpha at approximately 5-10 ng/ml, LFN gamma at approximately 20-50 ng/ml, IL-4 at approximately 5-10 ng/ml, IL-9 at approximately 5-10 ng/ml, IL-13 at approximately 5-10 ng/ml. Endothelial cells were sometimes starved for various times by culture in the basal media from Clonetics with 0.1 % semm.

Mononuclear cells were prepared from blood of employees at CuraGen Coφoration, using Ficoll. LAK cells were prepared from these cells by culture in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco/Life Technologies, Rockville, MD), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco) and Interleukin 2 for 4-6 days. Cells were then either activated with 10-20 ng/ml PMA and 1-2 μg/ml ionomycin, IL-12 at 5-10 ng/ml, LFN gamma at 20-50 ng/ml and IL-18 at 5-10 ng/ml for 6 hours. In some cases, mononuclear cells were cultured for 4-5 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco) with PHA (phytohemagglutinin) or PWM (pokeweed mitogen) at approximately 5 μg/ml. Samples were taken at 24, 48 and 72 hours for RNA preparation. MLR (mixed lymphocyte reaction) samples were obtained by taking blood from two donors, isolating the mononuclear cells using Ficoll and mixing the isolated mononuclear cells 1:1 at a final concentration of approximately 2xl0⁶ cells/ml in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol (5.5 x 10^"5 M) (Gibco), and 10 mM Hepes (Gibco). The MLR was cultured and samples taken at various time points ranging from 1- 7 days for RNA preparation.

Monocytes were isolated from mononuclear cells using CD 14 Miltenyi Beads, +ve VS selection columns and a Vario Magnet according to the manufacturer's instmctions. Monocytes were differentiated into dendritic cells by culture in DMEM 5% fetal calf semm (FCS) (Hyclone, Logan, UT), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco), 50 ng/ml GMCSF and 5 ng/ml IL-4 for 5-7 days. Macrophages were prepared by culture of monocytes for 5-7 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), 10 mM Hepes (Gibco) and 10% AB Human Semm or MCSF at approximately 50 ng/ml. Monocytes, macrophages and dendritic cells were stimulated for 6 and 12-14 hours with lipopolysaccharide (LPS) at 100 ng/ml. Dendritic cells were also stimulated with anti-CD40 monoclonal antibody (Pharmingen) at 10 μg/ml for 6 and 12-14 hours.

CD4 lymphocytes, CD8 lymphocytes and NK cells were also isolated from mononuclear cells using CD4, CD8 and CD56 Miltenyi beads, positive VS selection columns and a Vario Magnet according to the manufacturer's instmctions. CD45RA and CD45RO CD4 lymphocytes were isolated by depleting mononuclear cells of CD8, CD56, CD 14 and CD 19 cells using CD8, CD56, CD 14 and CD 19 Miltenyi beads and positive selection. CD45RO beads were then used to isolate the CD45RO CD4 lymphocytes with the remaining cells being CD45RA CD4 lymphocytes. CD45RA CD4, CD45RO CD4 and CD8 lymphocytes were placed in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco) and plated at 10⁶ cells/ml onto Falcon 6 well tissue culture plates that had been coated overnight with 0.5 μg/ml anti-CD28 (Pharmingen) and 3 ug/ml anti-CD3 (OKT3, ATCC) in PBS. After 6 and 24 hours, the cells were harvested for RNA preparation. To prepare chronically activated CD8 lymphocytes, we activated the isolated CD8 lymphocytes for 4 days on anti-CD28 and anti-CD3 coated plates and then harvested the cells and expanded them in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco) and IL-2. The expanded CD8 cells were then activated again with plate bound anti-CD3 and anti-CD28 for 4 days and expanded as before. RNA was isolated 6 and 24 hours after the second activation and after 4 days ofthe second expansion culture. The isolated NK cells were cultured in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco) and IL-2 for 4-6 days before RNA was prepared.

To obtain B cells, tonsils were procured from NDRI. The tonsil was cut up with sterile dissecting scissors and then passed through a sieve. Tonsil cells were then spun down and resupended at 10⁶ cells/ml in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco). To activate the cells, we used PWM at 5 μg/ml or anti-CD40 (Pharmingen) at approximately 10 μg/ml and IL-4 at 5-10 ng/ml. Cells were harvested for RNA preparation at 24,48 and 72 hours.

To prepare the primary and secondary Thl/Th2 and Trl cells, six-well Falcon plates were coated overnight with 10 μg/ml anti-CD28 (Pharmingen) and 2 μg/ml OKT3 (ATCC), and then washed twice with PBS. Umbilical cord blood CD4 lymphocytes (Poietic Systems,

5 6

German Town, MD) were cultured at 10 -10 cells/ml in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"

⁵ M (Gibco), 10 mM Hepes (Gibco) and IL-2 (4 ng/ml). IL-12 (5 ng/ml) and anti-IL4 (1 μg/ml) were used to direct to Thl, while IL-4 (5 ng/ml) and anti-LFN gamma (1 μg/ml) were used to direct to Th2 and IL-10 at 5 ng/ml was used to direct to Trl . After 4-5 days, the activated Thl, Th2 and Trl lymphocytes were washed once in DMEM and expanded for 4-7 days in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), 10 mM Hepes (Gibco) and IL-2 (1 ng/ml). Following this, the activated Thl, Th2 and Trl lymphocytes were re-stimulated for 5 days with anti-CD28/OKT3 and cytokines as described above, but with the addition of anti- CD95L (1 μg/ml) to prevent apoptosis. After 4-5 days, the Thl, Th2 and Trl lymphocytes were washed and then expanded again with IL-2 for 4-7 days. Activated Thl and Th2 lymphocytes were maintained in this way for a maximum of three cycles. RNA was prepared from primary and secondary Thl, Th2 and Trl after 6 and 24 hours following the second and third activations with plate bound anti-CD3 and anti-CD28 mAbs and 4 days into the second and third expansion cultures in Interleukin 2.

The following leukocyte cells lines were obtained from the ATCC: Ramos, EOL-1, KU-812. EOL cells were further differentiated by culture in 0.1 mM dbcAMP at 5 xlO⁵ cells/ml for 8 days, changing the media every 3 days and adjusting the cell concentration to 5 xlO⁵ cells/ml. For the culture of these cells, we used DMEM or RPMI (as recommended by the ATCC), with the addition of 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), 10 mM Hepes (Gibco). RNA was either prepared from resting cells or cells activated with PMA at 10 ng/ml and ionomycin at 1 μg/ml for 6 and 14 hours. Keratinocyte line CCD 106 and an airway epithelial tumor line NCI-H292 were also obtained from the ATCC. Both were cultured in DMEM 5% FCS (Hyclone), 100 μM non essential amino acids (Gibco), 1 mM sodium pyruvate (Gibco), mercaptoethanol 5.5 x 10^"5 M (Gibco), and 10 mM Hepes (Gibco). CCD 1106 cells were activated for 6 and 14 hours with approximately 5 ng/ml TNF alpha and 1 ng/ml IL-1 beta, while NCI-H292 cells were activated for 6 and 14 hours with the following cytokines: 5 ng/ml IL-4, 5 ng/ml IL-9, 5 ng/ml IL-13 and 25 ng/ml LFN gamma.

For these cell lines and blood cells, RNA was prepared by lysing approximately 10⁷ cells/ml using Trizol (Gibco BRL). Briefly, 1/10 volume of bromochloropropane (Molecular Research Coφoration) was added to the RNA sample, vortexed and after 10 minutes at room temperature, the tubes were spun at 14,000 φm in a Sorvall SS34 rotor. The aqueous phase was removed and placed in a 15 ml Falcon Tube. An equal volume of isopropanol was added and left at -20 degrees C overnight. The precipitated RNA was spun down at 9,000 φm for 1 min in a Sorvall SS34 rotor and washed in 70% ethanol. The pellet was redissolved in 300 μl of RNAse-free water and 35 μl buffer (Promega) 5 μl DTT, 7 μl RNAsin and 8 μl DNAse were added. The tube was incubated at 37 degrees C for 30 minutes to remove contaminating genomic DNA, extracted once with phenol chloroform and re-precipitated with 1/10 volume of 3 M sodium acetate and 2 volumes of 100% ethanol. The RNA was spun down and placed in RNAse free water. RNA was stored at -80 degrees C.

Al comprehensive panel vl.O The plates for AI_comprehensive panel l.O include two control wells and 89 test samples comprised of cDNA isolated from surgical and postmortem human tissues obtained from the Backus Hospital and Clinomics (Frederick, MD). Total RNA was extracted from tissue samples from the Backus Hospital in the Facility at CuraGen. Total RNA from other tissues was obtained from Clinomics.

Joint tissues including synovial fluid, synovium, bone and cartilage were obtained from patients undergoing total knee or hip replacement surgery at the Backus Hospital. Tissue samples were immediately snap frozen in liquid nitrogen to ensure that isolated RNA was of optimal quality and not degraded. Additional samples of osteoarthritis and rheumatoid arthritis joint tissues were obtained from Clinomics. Normal control tissues were supplied by Clinomics and were obtained during autopsy of trauma victims.

Surgical specimens of psoriatic tissues and adjacent matched tissues were provided as total RNA by Clinomics. Two male and two female patients were selected between the ages of 25 and 47. None ofthe patients were taking prescription dmgs at the time samples were isolated.

Surgical specimens of diseased colon from patients with ulcerative colitis and Crohns disease and adjacent matched tissues were obtained from Clinomics. Bowel tissue from three female and three male Crohn's patients between the ages of 41-69 were used. Two patients, were not on prescription medication while the others were taking dexamethasone, phenobarbital, or tylenol. Ulcerative colitis tissue was from three male and four female patients. Four ofthe patients were taking lebvid and two were on phenobarbital.

Total RNA from post mortem lung tissue from trauma victims with no disease or with emphysema, asthma or COPD was purchased from Clinomics. Emphysema patients ranged in age from 40-70 and all were smokers, this age range was chosen to focus on patients with cigarette-linked emphysema and to avoid those patients with alpha- lanti-trypsin deficiencies. Asthma patients ranged in age from 36-75, and excluded smokers to prevent those patients that could also have COPD. COPD patients ranged in age from 35-80 and included both smokers and non-smokers. Most patients were taking corticosteroids, and bronchodilators.

In the labels employed to identify tissues in the AI_comprehensive panel_vl.O panel, the following abbreviations are used:

AI = Autoimmunity Syn = Synovial Normal = No apparent disease Rep22 /Rep20 = individual patients RA = Rheumatoid arthritis

Backus = From Backus Hospital OA = Osteoarthritis (SS) (BA) (MF) = Individual patients Adj = Adjacent tissue Match control = adjacent tissues -M = Male

-F = Female COPD = Chronic obstmctive pulmonary disease

Panels 5D and 51 The plates for Panel 5D and 51 include two control wells and a variety of cDNAs isolated from human tissues and cell lines with an emphasis on metabolic diseases. Metabolic tissues were obtained from patients enrolled in the Gestational Diabetes study. Cells were obtained during different stages in the differentiation of adipocytes from human mesenchymal stem cells. Human pancreatic islets were also obtained. In the Gestational Diabetes study subjects are young (18 - 40 years), otherwise healthy women with and without gestational diabetes undergoing routine (elective) Caesarean section. After delivery ofthe infant, when the surgical incisions were being repaired/closed, the obstetrician removed a small sample (<1 cc) ofthe exposed metabolic tissues during the closure of each surgical level. The biopsy material was rinsed in sterile saline, blotted and fast frozen within 5 minutes from the time of removal. The tissue was then flash frozen in liquid nitrogen and stored, individually, in sterile screw-top tubes and kept on dry ice for shipment to or to be picked up by CuraGen. The metabolic tissues of interest include uterine wall (smooth muscle), visceral adipose, skeletal muscle (rectus) and subcutaneous adipose. Patient descriptions are as follows: Patient 2 Diabetic Hispanic, overweight, not on insulin

Patient 7-9 Nondiabetic Caucasian and obese (BMI>30)

Patient 10 Diabetic Hispanic, overweight, on insulin

Patient 11 Nondiabetic African American and overweight

Patient 12 Diabetic Hispanic on insulin

Adipocyte differentiation was induced in donor progenitor cells obtained from Osims

(a division of Clonetics/BioWhittaker) in triplicate, except for Donor 3U which had only two replicates. Scientists at Clonetics isolated, grew and differentiated human mesenchymal stem cells (HuMSCs) for CuraGen based on the published protocol found in Mark F. Pittenger, et al., Multilineage Potential of Adult Human Mesenchymal Stem Cells Science Apr 2 1999:

143-147. Clonetics provided Trizol lysates or frozen pellets suitable for mRNA isolation and ds cDNA production. A general description of each donor is as follows:

Donor 2 and 3 U Mesenchymal Stem cells Undifferentiated Adipose Donor 2 and 3 AM Adipose AdiposeMidway Differentiated

Donor 2 and 3 AD Adipose Adipose Differentiated

Human cell lines were generally obtained from ATCC (American Type Culture Collection), NCI or the German tumor cell bank and fall into the following tissue groups: kidney proximal convoluted tubule, uterine smooth muscle cells, small intestine, liver HepG2 cancer cells, heart primary stromal cells, and adrenal cortical adenoma cells. These cells are all cultured under standard recommended conditions and RNA extracted using the standard procedures. All samples were processed at CuraGen to produce single stranded cDNA. Panel 51 contains all samples previously described with the addition of pancreatic islets from a 58 year old female patient obtained from the Diabetes Research Institute at the University of Miami School of Medicine. Islet tissue was processed to total RNA at an outside source and delivered to CuraGen for addition to panel 51.

In the labels employed to identify tissues in the 5D and 51 panels, the following abbreviations are used:

GO Adipose = Greater Omentum Adipose

SK = Skeletal Muscle UT = Utems PL = Placenta AD = Adipose Differentiated

AM = Adipose Midway Differentiated U = Undifferentiated Stem Cells

Panel CNSD.01 The plates for Panel CNSD.01 include two control wells and 94 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center. Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at -80°C in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.

Disease diagnoses are taken from patient records. The panel contains two brains from each ofthe following diagnoses: Alzheimer's disease, Parkinson's disease, Huntington's disease, Progressive Supernuclear Palsy, Depression, and "Normal controls". Within each of these brains, the following regions are represented: cingulate gyms, temporal pole, globus palladus, substantia nigra, Brodman Area 4 (primary motor strip), Brodman Area 7 (parietal cortex), Brodman Area 9 (prefrontal cortex), and Brodman area 17 (occipital cortex). Not all brain regions are represented in all cases; e.g., Huntington's disease is characterized in part by neurodegeneration in the globus palladus, thus this region is impossible to obtain from confirmed Huntington's cases. Likewise Parkinson's disease is characterized by degeneration ofthe substantia nigra making this region more difficult to obtain. Normal control brains were examined for neuropathology and found to be free of any pathology consistent with neurodegeneration.

In the labels employed to identify tissues in the CNS panel, the following abbreviations are used:

PSP = Progressive supranuclear palsy Sub Nigra = Substantia nigra Glob Palladus= Globus palladus

Temp Pole = Temporal pole Cing Gyr = Cingulate gyms BA 4 = Brodman Area 4

Panel CNS_Neurodegeneration_V1.0

The plates for Panel CNS_Neurodegeneration_V1.0 include two control wells and 47 test samples comprised of cDNA isolated from postmortem human brain tissue obtained from the Harvard Brain Tissue Resource Center (McLean Hospital) and the Human Brain and Spinal Fluid Resource Center (VA Greater Los Angeles Healthcare System). Brains are removed from calvaria of donors between 4 and 24 hours after death, sectioned by neuroanatomists, and frozen at -80°C in liquid nitrogen vapor. All brains are sectioned and examined by neuropathologists to confirm diagnoses with clear associated neuropathology.

Disease diagnoses are taken from patient records. The panel contains six brains from Alzheimer's disease (AD) patients, and eight brains from "Normal controls" who showed no evidence of dementia prior to death. The eight normal control brains are divided into two categories: Controls with no dementia and no Alzheimer's like pathology (Controls) and controls with no dementia but evidence of severe Alzheimer's like pathology, (specifically senile plaque load rated as level 3 on a scale of 0-3; 0 = no evidence of plaques, 3 = severe AD senile plaque load). Within each of these brains, the following regions are represented: hippocampus, temporal cortex (Brodman Area 21), parietal cortex (Brodman area 7), and occipital cortex (Brodman area 17). These regions were chosen to encompass all levels of neurodegeneration in AD. The hippocampus is a region of early and severe neuronal loss in AD; the temporal cortex is known to show neurodegeneration in AD after the hippocampus; the parietal cortex shows moderate neuronal death in the late stages ofthe disease; the occipital cortex is spared in AD and therefore acts as a "control" region within AD patients. Not all brain regions are represented in all cases. In the labels employed to identify tissues in the CNS_Neurodegeneration_V1.0 panel, the following abbreviations are used:

AD = Alzheimer's disease brain; patient was demented and showed AD-like pathology upon autopsy

Control = Control brains; patient not demented, showing no neuropathology

Control (Path) = Control brains; pateint not demented but showing sever AD-like pathology

SupTemporal Ctx = Superior Temporal Cortex

Inf Temporal Ctx = Inferior Temporal Cortex

NOVla: Membrane Protein/neuropilin/metalloproteinase-like

Expression ofthe NOVla gene (SC40376139) was assessed using the primer-probe set Ag2229 described in Table 11. Results from RTQ-PCR mns are shown in Tables 12 and 13.

Table 11. Probe Name Ag2229

Table 12. Panel 1.3D

Table 13. Panel CNS_neurodegeneration_vl.O

Panel 1.3D Summary Expression ofthe NOV1 gene appears to be brain-specific. Highest expression is detected in the hippocampus (CT=33) and the cerebral cortex, two regions that degenerate in Alzheimer's disease. Thus, this gene would be useful for distinguishing brain tissue from non-neural tissue, and may be beneficial as a dmg target in neurodegenerative disease. Please see Panel_CNS_neurodegeneration for further discussion of potential utility in the central nervous system.

Panel 2D Summary Expression ofthe NOV1 gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.) Panel 4D Summary Expression ofthe NOV1 gene is low/undetectable (Ct values

>35) in all samples in this panel. (Data not shown.)

Panel CNS_1 Summary Expression ofthe NOV1 gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.)

Panel CNS neurodegeneration vl.O Summary The NOV1 gene is expressed at low but significant levels in the brain, as is seen in Panel 1.3D, with highest expression in the occipital cortex from the brain of a control patient (CT=32.2). Importantly, once the CT values are corrected for RNA quality/amount per well, there appears to be a down-regulation in the temporal cortex ofthe Alzheimer's diseased (AD) brain (in 5 of 6 diseased brains), a region that is known to undergo severe neurodegeneration. Conversely, in the occipital cortex, where neurodegeneration is not observed in AD, this gene does not appear to be differentially regulated. Neuropilins act as receptors for semaphorins, which mediate axon repulsion during neurodevelopment as well as neuroapoptosis. Thus, the downregulation ofthe expression of this gene may be critically involved in the neurodegeneration seen in Alzheimer's disease.

Therefore, selective antagonism of this receptor may slow neurodegeneration in this disease or enhance the compensatory synaptogenesis process. Furthermore, the NOVl gene product may be useful in any clinical situation involving neuronal death, such as Parkinson's or Huntington's disease, spinocerebellar ataxia, head or spinal cord trauma, or stroke (Zhang et al., Up-regulation of neuropilin-1 in neovasculature after focal cerebral ischemia in the adult rat. J Cereb Blood Flow Metab. 21(5): 541-9, 2001; Ziv et al., Semaphorins as mediators of neuronal apoptosis. J Neurochem. 73(3):961-71, 1999). NOV2: Fibrillin-like protein

Expression ofthe NOV2a gene (GMAC022146_A) and variants NOV2b (153568997), NOV2c (CG88987-01), NOV2d (CG88987-02), NOV2e (CG88987-03) and NOV2f (CG88987-05) was assessed using the primer-probe sets Ag72, Ag390, Ag671, Ag766, Agl211, Agl928, Agl938, Ag2233, Ag2241, Ag4154, and Ag4334 described in Tables 11- 24. Please note that not all probe and primer sets match all the sequences. Table 25 provides a complete summary ofthe probe and primer sets that correspond to each sequence, where the "X" represents a match between the sequence and the probe and primer set. Results from RTQ-PCR mns are shown in Tables 26-35.

Table 14. Probe Name Ag72

Table 15. Probe Name Ag390

Table 17. Probe Name Aε766

Start SEQ ID

Primers Sequences TM Length Position NO:

Forward 5 ' -TGCTACTACCTGCCTGGATATG-3 ' 59.3 22 839 140

Table 25.

Table 26. Panel 1

Table 27. Panel 1.1

Table 28. Panel 1.2

Table 29. Panel 1.3D

Table 30. Panel General_screening_panel_vl.4

Table 31. Panel 2.2

Table 32. Panel 2D

Table 34. Panel 4. ID

Panel 1 Summary Ag72/Ag390 Multiple runs with two different probe and primer sets show highest expression ofthe NOV2 gene in the fetal brain (CTs=25-27), fetal lung and a cell line derived from a gastric cancer. Thus, the expression of this gene could be used to distinguish fetal brain tissue from adult brain tissue and fetal lung from adult lung.

Significant expression is also seen in fetal kidney, when compared to expression in the adult kidney. Therefore, the expression of this gene could also be used to distinguish fetal kidney tissue from adult kidney tissue.

Panel 1.1 Summary Ag671 Highest expression ofthe NOV2 gene is seen in the fetal brain (CT=26.2), a result that is repeated in Panel 1. Significant expression is also seen in fetal lung and skeletal muscle (CTs=26.7-32.8) when compared to expression in the adult tissues. In addtion, low, but significant expression is seen in the gastric cancer cell line (NCI-N87) and in adult and fetal kidney. Thus, the expression of this gene could be used to distinguish fetal brain tissue from other tissues in the panel. In addition, the expression of this gene could be used to distinguish fetal lung and skeletal mucle tissue from adult lung and skeletal muscle tissue.

Panel 1.2 Summary Ag766 Highest expression ofthe NOV2 gene in this panel is in the fetal brain (CT=27). Low, but significant expression is also seen in other brain samples including the amygdala, hippocampus, thalamus, and cerebral cortex. The higher levels of expression present in the fetal brain when compared to expression in the adult brain suggest an ongoing role in CNS processes.

There is also substantial expression in samples derived from a gastric cancer cell line (NCI-N87), fetal lung tissue and adult and fetal kidney tissue. This expression profile is concordant with the expression seen in Panels 1 , and 1.1. Of note is the difference in expression between fetal lung (CT=27.7) and its adult counterpart (CT=31.8). Thus, the expression of this gene could be used to distinguish fetal lung tissue from adult lung tissue.

Panel 1.3D Summary Agl211/Ag2233 Three runs with two different probe and primer sets show highest expression in the fetal brain, lung and skeletal muscle (CTs=30). There is also substantial expression in samples derived from a gastric cancer cell line (NCI- N87). Thus, the expression of this gene could be used to distinguish fetal skeletal muscle or fetal lung from their respective adult tissue counterparts.

Ag2241 Expression ofthe NOV2 gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.) Panel General_screening_panel_vl.O Summary Agl928/Ag4154/Ag4334 Multiple runs with different probe and primer sets show highest expression ofthe NOV2 gene in the fetal brain (CTs=26-33), indicating a probable developmental role for the NOV2 gene in the CNS. This finding is consistent with the expression seen in Panels 1, 1.1, 1.2 and 1.3D. There is also significant expression in the fetal kidney and lung and in the gastric cancer cell line (NCI-N87). Thus, the expression of this gene could be used to distinguish fetal brain tissue from adult brain. In addition, the expression of this gene could be used to distinguish fetal kidney tissue from adult kidney tissue and fetal lung tissue from adult lung tissue.

Panel 2.2. Summary Ag2241 The expression ofthe NOV2 gene appears to be exclusive to a sample derived from a kidney cancer. Thus, the expression of this gene could be used to distinguish this sample from others in the panel, including its normal adjacent tissue. Moreover, therapeutic modulation of this gene or its protein product, through the use of small molecule drugs, antibodies or protein therapeutics, may be useful in the treatment of kidney cancer.

Panel 2D Summary Agl211 Two runs using the same probe and primer produce results that are in excellent agreement, with highest expression ofthe NOV2 gene in a kidney cancer (CTs=29.5). This expression profile is in concordance with the expression seen in Panel 2.2. Significant expression is also seen in a thyroid cancer sample and a series of normal adjacent tissues derived from kidney and lung cancer cases. Thus, the expression of this gene could be used to distinguish this kidney cancer sample from others in the panel, including its normal adjacent tissue. In addition, low levels of expression could be used to distinguish normal adjacent tissue from adjacent malignant tissue. Moreover, therapeutic modulation of this gene or its protein product, through the use of small molecule drugs, antibodies or protein therapeutics, might be of benefit in the treatment of kidney cancer. Panel 4D Summary Agl938/Ag2233 Expression of the NOV2 gene is limited to a few samples in this panel, with highest expression seen in the thymus. This expression is consistent in two runs with two different probe and primer sets (CTs= 33-34). Significant expression is also seen in the normal lung, a result that is replicated in Panel 4. ID and Panels 1, 1.2, and 1.3D. Please see Panel 4. ID for potential utility of this gene in the treatment of autoinflammatory related disease.

Agl928/Ag2241 Expression ofthe NOV2 gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.)

Panel 4.1 D Summary Ag4154/Ag4334 Two experiments with two different probes and primers both show highest expression ofthe NOV2 gene in the kidney (CTs=33-34). There is also low but significant expression in normal lung and thymus. The expression in normal kidney is consistent with the expression seen in Panels 1, 1.1, 1.2, 1.3D and Panel 4. The expression of low levels ofthe NOV2 gene product, a secreted fibrillin-related homolog, in the normal lung, thymus, and kidney suggest that it could be used as a protein or antibody therapeutic to reduce or eliminate symptoms of patients with connective tissue diseases ofthe lung, thymus and kidney. These diseases include Marfan syndrome, ankylosing spondylitis, Sjogren's syndrome, and relapsing polychondritis.

Panel CNS_neurodegeneration_vl.O SummaryAg4154/Ag4334Expression ofthe NOV2 gene is restricted to a few samples with highest expression in the cerebral cortex of a control patient. This gene does not appear to be expressed in the brains of patients with Alzheimer's disease. Please see Panel 1 for discussion of potential utility in the CNS.

Ag2241 Expression ofthe NOV2 gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.)(Tanoue, Pulmonary involvement in collagen vascular disease: a review ofthe pulmonary manifestations ofthe Marfan syndrome, ankylosing spondylitis, Sjogren's syndrome, and relapsing polychondritis. 7:62-77, 1992; Kanwar et al., Isolation of rat fibrillin-1 cDNA and its relevance in metanephric development. 275 (5 Pt 2):F710-23, 1998).

NOV3: KIAAl 589-like

Expression ofthe NOV3 gene (GSAL442663.1_A) was assessed using the primer- probe set Agl550 described in Table 36. Results from RTQ-PCR runs are shown in Table 37. Table 36. Probe Name Agl550

Table 37. Panel 1.3D

Panel 1.3D Summary The NOV3 gene, a KIAAl 589 homolog, is ubiquitously expressed in this panel, with highest expression seen in fetal skeletal muscle (CT=29). Expression ofthe gene appears to be much higher in fetal skeletal muscle than in adult skeletal muscle (CT=32.6). In addition, expression ofthe gene appears to be higher in fetal heart (CT=30.5) than in adult heart (CT=33.9). Thus, expression of this gene could be used to distinguish between adult and fetal sources of heart and skeletal muscle. In addition, the higher levels of expression ofthe NOV3 gene in fetal heart and skeletal muscle when compared to the levels of expression in adult tissue, suggests that the protein encoded by the NOV3 gene may be involved in the development of these tissues. Therefore, therapeutic modulation of this gene or its protein product may be effective in the treatment of diseases that affect the heart, such as atherosclerosis, hypertension, or aortic stenosis. Furthermore, the therapeutic modulation of this gene or gene product, through replacement therapy, could be used as a regenerative therapy for muscle disease. Among tissues involved in central nervous system function, this gene is expressed at moderate levels in all brain regions examined. The NOV3 gene encodes a protein with a putative zinc-finger motif. Since these proteins are known to interact with nucleic acids, this suggests that the NOV3 gene product may play a potential role in transcription. Thus, therapeutic modulation ofthe NOV3 gene product may be used to regulate the transcription of disease-related proteins such as ataxin, huntingtin, or various apoptosis cascade proteins.

NOV4: WD40 motif protein Expression ofthe NOV4 gene (GSAL442663.1 B) was assessed using the primer- probe sets Agl551 and Ag3362 described in Tables 38 and 39. Results from RTQ-PCR runs are shown in Tables 40-44.

Table 38. Probe Name Agl551

Table 39. Probe Name Ag3362

Table 40. Panel 1.3D

Table 41. Panel General_screening_panel_vl.O

Table 42. Panel 2.2

Table 43. Panel 4D

Table 44. Panel CNS_neurodegeneration_V1.0

Panel 1.3D Summary Agl551 The NOV4 gene is widely expressed in this panel, with highest expression in the breast cancer cell line MCF-7 (CT=29). Of note is the difference in expression between the fetal and adult heart and skeletal muscle, with higher expression seen in the tissues derived from the fetal source (CTs=30-32) than in tissues derived from the adult (CTs=34-36). Thus, expression ofthe NOV4 gene could be used to distinguish between fetal and adult heart and skeletal muscle. In general, expression ofthe NOV4 gene appears to be greater in the cancer cell lines and fetal cells than in the samples derived from normal tissues. Since normal cultured cell lines and fetal cells are highly proliferative, this observation may indicate that the expression ofthe NOV4 gene might be used to distinguish proliferating cells over resting or quiescent cells.

Among tissues involved in central nervous system function, this gene is expressed at moderate levels in all brain regions examined. The NOV4 gene encodes a protein with a putative WD40 motif, which is known to interact with the G protein beta subunit (Gbeta), suggesting a role in signal transduction. Several neurotransmitter receptors are GPCRs, including the dopamine receptor family, the serotonin receptor family, the GABAB receptor, muscarinic acetylcholine receptors, and others; thus this molecule may mediate signaling via a neurotransmitter receptor. Targeting various neurotransmitter receptors (dopamine, serotonin) has proven to be an effective therapy in psychiatric illnesses such as schizophrenia, bipolar disorder and depression. Therefore, therapeutic modulation of this gene or its protein product may be beneficial in one or more of these diseases, as may antagonism ofthe protein encoded by the gene. Among tissues with metabolic function, the NOV4 gene is expressed in adipose, adrenal gland, adult and fetal heart, adult and fetal liver, pancreas, pituitary and thyroid. This putative nuclear protein may be important for the pathogenesis and/or treatment of disease in any or all of these tissues, including obesity and diabetes.

Panel General_screening_panel_vl.O Summary Ag3362 The NOV4 gene is widely expressed in this panel, with highest expression in the breast cancer cell line T47D (CT=29). Significant expression is also seen in cell lines derived from prostate, breast and ovarian cancers. In general, expression ofthe NOV4 gene appears to be greater in the cancer cell lines than in normal tissue, an observation that is consistent with the results from Panel 1.3D. Thus, the expression of this gene could be used to distinguish these cell line types from others in the panel.

Panel 2.2 Summary Agl551 Highest expression ofthe NOV4 gene is seen in normal kidney tissue adjacent to malignant kidney tissue. Thus, the expression of this gene could be used to distinguish this sample from others in the panel.

Panel 4D Summary Agl551 The NOV4 gene is widely expressed in this panel, a pattern detected in the other panels as well, with highest expression in the ionomycin activated B cell line (Ramos) (CT=28.6). Significant expression is also detected in normal B lymphocytes, a B cell line (Ramos), and in normal B cells activated by pokeweed mitogen. Cytoplasmic and nuclear localization (PSORT) suggests that this G protein-beta-WD40 motif protein homolog may be used as a target for small molecule drug discovery. Activated B lymphocytes are important antigen presenting cells that participate in the stimulation ofthe immune response in numerous settings. Inhibitory small molecule drugs that reduce the function of activated B cells may reduce or eliminate symptoms in patients with autoimmune and inflammatory diseases such as lupus erythematosus, Crohn's disease, ulcerative colitis, asthma, psoriasis, and rheumatoid arthritis. Please note that data from one run with the probe and primer set Ag3362 is not included because the amp plot corresponding to the run indicates that there were problems with the experiment.

Panel CNS_neurodegeneration_V1.0 Summary Agl551/Ag336 Highest expression ofthe the NOV4 gene is seen in the occipital cortex of a control patient and the temporal cortex of an Alzheimer's patient. While the NOV4 gene does not appear to be preferentially expressed in Alzheimer's disease, this panel confirms expression ofthe NOV4 gene at moderate/high levels in the brain in an additional set of individuals. Please see Panel 1.3D for discussion of potential utility of this gene in the central nervous system (Zhu et al., Transcription activating property of autoantigen SG2NA and modulating effect of WD-40 repeats. Exp Cell Res. 269(2):312-21, 2001).

NOV5a and NOV5b: Novel Opioid Binding Cell Adhesion Molecule

Expression ofthe NOV5a gene (139785504) and the NOV5b variant (139785504_dal) was assessed using the primer-probe sets Ag3090 and Ag3092 described in Tables 45 and 46. Results from RTQ-PCR runs are shown in Tables 47-50.

Table 45. Probe Name Ag3090

Table 47. Panel 1.3D

Table 48. Panel 4D

Table 49. Panel CNS 1

Panel 1.3D Summary Ag3090/Ag3092 Two experiments with two different probe and primer sets produce results that are in very good agreement, with highest expression ofthe NOV5A gene in regions ofthe brain, specifically the thalamus and fetal brain (CTS=29-30). Indeed, the NOV5A gene shows a highly brain-preferential expression, and is expressed at high levels in all brain regions examined. The protein encoded by the NOV5A gene is a homolog of opioid- binding cell adhesion molecule (OBCAM), which is believed to be involved in axonal outgrowth. In addition, this molecule may be involved in the synaptic machinery. The limited ability ofthe CNS to regenerate after injury is the major impediment to the treatment of stroke, spinal cord and head trauma, and neurodegenerative diseases. Therefore, selective modulation of this gene or its protein product may be useful in enhancing neuroregeneration in any or all of these clinical conditions.

Panel 2.2 Summary Ag3090/Ag3092 Expression ofthe NOV5A gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.) Panel 4D Summary Ag3092 Expression of the NOV5 A gene is limited to a sample derived from secondary Th2 cells (Ct=34.2). This expression profile suggests that the NOV5A gene product may be useful as a protein therapeutic or a target for the generation of therapeutic antibodies that reduce or eliminate the symptoms in patients with one or more diseases related to the functions of Th2 cells, including asthma and allergies. Ag3090 Expression of the NOV5 A gene is low/undetectable (Ct values >35) in all samples in this panel. (Data not shown.)

Panel CNS_1 Summary Ag3092 Highest expression ofthe NOV5A gene is seen in the parietal cortex (Brodman's Area 7) of a control patient (CT=31.2). The gene is also widely expressed across many regions ofthe brain in many disease states. Please see Panel 1.3D for discussion of potential utility in the central nervous system.

Panel CNS_neurodegeneration_V1.0 Ag3090/Ag3092 Two experiments with two different probe and primer sets produce results that are in very good agreement, with highest expression in the occipital cortex and the parietal cortex of a control patient (CT=30.3). While the expression of this gene does not appear to be specific to Alzheimer's disease, the results of this panel confirm expression ofthe NOV5A gene at moderate/high level in the brain in an independent set of individuals. Please see Panel 1.3D for a discussion of potential utility in the central nervous system. (Hachisuka et al., Localization of opioid-binding cell adhesion molecule (OBCAM) in adult rat brain. Brain Res. 842(2):482-6, 1999; Hachisuka et al., Developmental expression of opioid-binding cell adhesion molecule (OBCAM) in rat brain. Brain Res Dev Brain Res 122(2):183-91, 2000).

NOV5d: Opioid Binding Cell Adhesion Molecule

Expression ofthe NOV5d gene (CG51027-05) was assessed using the primer-probe set Ag565 described in Table 51. Results from RTQ-PCR runs are shown in Tables 52-54.

Table 51. Probe Name Ag565

ble 52. Panel 1.1

Table 53. Panel 4. ID

Panel 1.1 Summary The NOV5D gene is expressed most highly in the cerebellum (CT=22). Indeed, this gene shows a highly brain-preferential expression, and is expressed at high levels in all brain regions examined. The protein encoded by the NOV5D gene is a homolog of opioid-binding cell adhesion molecule (OBCAM), which is believed to be involved in axonal outgrowth. In addition, this molecule may be involved in the synaptic machinery. The limited ability ofthe CNS to regenerate after injury is the major impediment to treatment of stroke, spinal cord and head trauma, and neurodegenerative diseases. Therefore, selective modulation of this gene or its protein product may be useful in enhancing neuroregeneration in any or all of these clinical conditions.

There is also significant expression in tissue derived from fetal heart and kidney (CTs=26-29) when compared to the level of expression in the adult source of these tissues (CTs=32-37). Thus, expression of this gene could be used to differentiate between the adult and fetal sources of heart and kidney tissue.

The NOV5D gene encodes a putative intracellular protein with moderate expression in many metabolic tissues including adrenal, adult and fetal liver, pancreas, pituitary and thyroid. Therefore, this protein may be important for the pathogenesis and/or treatment of disease in any or all of these tissues, including obesity and diabetes.

Panel General_screening_panel_vl.4 Summary Please note that data from this experiment is not included because the amp plot corresponding to the run indicates that there were problems with the run.

Panel 4.1D Summary Expression is limited to a few samples in this panel, with highest expression in the kidney (CT=30.4) and thymus. Moderate expression is seen dibutyryl -cAMP-differentiated and phorbol ester plus ionomycin-activated eosinophil cell line EOL-1, with low but significant levels of expression detected in unstimulated astrocytes. This expression profile suggests that the NOV5D gene product, an opioid-binding cell adhesion molecule homolog, may be useful as a therapeutic protein to reduce or eliminate symptoms resulting from diseases ofthe thymus and kidney. Furthermore, the protein encoded by the NOV5D gene may also reduce or eliminate allergies in which activated eosinophils play a role and may be effective in the treatment of multiple sclerosis where astrocytes present antigens to T lymphocytes.

Panel CNS neurodegeneration vl.O Summary The NOV5D gene is widely expressed in this panel, with highest expression detected in the occipital cortex of a control brain (CT=29.1). The expression seen across all regions ofthe brain is in agreement with the expression seen in Panel 1.1. While the expression of this gene does not appear be specific to Alzheimer's disease, this panel confirms expression at the moderate/high level in the brain in an additional set of individuals. Please see Panel 1.1 for discussion of potential utility in the central nervous system (Hachisuka et al., Localization of opioid-binding cell adhesion molecule (OBCAM) in adult rat brain. Brain Res. 842(2):482-6, 1999; Hachisuka et al., Developmental expression of opioid-binding cell adhesion molecule (OBCAM) in rat brain. Brain Res Dev Brain Res 122(2):183-91, 2000).

NOV6: Triacylglycerol lipase-like

Expression ofthe NOVόa gene (SCI 22982104_A) and the NOV6b variant (CG58608- 02) was assessed using the primer-probe sets Ag2179 and Ag3927 described in Tables 55 and 56. Please note that only the probe and primer set Ag3927 match the CG58608-02 sequence. Results from RTQ-PCR runs are shown in Table 57.

Table 55. Probe Name Ag2179

TET-5 ' -AAGACTTCTTGCCTAAAACCTCATTTAAAA-

Probe 63 . 7 30 761 183 TAMRA

Reverse 5 ' -ACACAGCTTTGAACCAATGAAT-3 ' 58 . 6 22 792 184

Table 56. Probe Name Ag3927

Start SEQ ID

Primers Sequences TM Length Position NO:

Forward 5 ' -GTCCTGATCTGAACTTGGTTCA-3 ' 59.2 22 980 185

TET-5 ' -CAGACAACGTCTCCATTATACAACATGACA-3 '

Probe 66.9 30 1009 186 TAMRA iReverse 5 ' -TTACCATTCCAAATTGCAGTTG-3 ' 59.9 22 1052 187

CNS ca. (astro) SNB-75 0.0

CNS ca. (glio) SNB-19 0.0

CNS ca. (glio) SF-295 2.8

113447 Brain (Amygdala) Pool 7.7

103382 Brain (cerebellum) 3.7

64019-1 brain(fetal) 27.5

113448 Brain (Hippocampus) Pool 0.0

113464 Cerebral Cortex Pool 9.6

113449_Brain (Substantia nigra) Pool 2.4

113450 Brain (Thalamus) Pool 9.0 103384 Brain (whole) 1.2

113458 Spinal Cord Pool 3.0

103375 Adrenal Gland 0.0

113454 Pituitary gland Pool 0.0

103397 Salivary Gland 0.9

103369 Thyroid (female) 24.7

Pancreatic ca. CAPAN2 1.4

113453 Pancreas Pool 1.8

Panel General_screening_panel_vl.4 Summary Ag3927 Expression ofthe NOV6a gene in this panel is limited to samples originating from the testis (CT=33) and the kidney. Thus, expression of this gene could be used to distinguish testis and kidney tissue from the other tissues in this panel.

Ag2179 Expression ofthe NOV6a gene is low/undetectable (Ct values >35) in all samples in Panels 1.3D, 2D, 4D, and CNS_neurodegeneration_vl.O. (Data not shown.)

NOV7a: IGE Receptor Beta Subunit

Expression ofthe NOV7a gene (SC126624027_A) was assessed using the primer- probe set Ag2178 described in Table 58. Results from RTQ-PCR runs are shown in Table 59.

Table 58. Probe Name Ag2178

Table 59. Panel 1.3D

Panel 1.3D Summary Expression ofthe NOV7a gene is restricted to the testis (CT=32.9). Thus, the expression of this gene could be used to distinguish testis tissue from the other samples on the panel.

Expression ofthe NOV7a gene is low/undetectable (Ct values >35) in all samples in Panels 2D and 4D. (Data not shown.)

NOV8: Muncl 8-like

Expression ofthe NOV8 gene (SC138745558_A) was assessed using the primer-probe set Ag2177 described in Table 60. Results from RTQ-PCR runs are shown in Tables 61-64.

Table 60. Probe Name Ag2177

Table 61. Panel 1.3D

Table 62. Panel 2D

Ta le 63. Panel 4D

. Panel 1.3D Summary Expression ofthe NOV8 gene is ubiquitous among the samples in this panel, with highest expression in a sample derived from a breast cancer cell line (BT- 549) (CT=27.4). In addition, there is substantial expression in a neuroblastoma cell line (SK- N-AS). Of note is the difference in expression between the sample of fetal skeletal muscle and its adult counterpart. Thus, the expression of this gene could be used to distinguish skeletal muscle from fetal and adult sources.

In addition, there is substantial expression of this gene in the hippocampus and cerebral cortex. Munc 18 appears to be critically involved in the process of synaptic vesicle docking prior to neurotransmitter release. Since the NOV8 gene product is a Munc 18 homolog, inhibition ofthe function of this protein may have therapeutic benefit in any disease in which selective decreases of neurotransmission has been shown to ameliorate symptomology (e.g., epilepsy or other seizure disorders, schizophrenia, bipolar disorder or anxiety).

The NOV8 gene encodes a putative intracellular protein with moderate expression in many metabolic tissues including adipose, adrenal gland, adult and fetal heart, adult and fetal liver, adult and fetal skeletal muscle, pancreas, pituitary and thyroid. Thus, this protein may be important for the pathogenesis and/or treatment of disease in any or all of these tissues, including obesity and diabetes.

Panel 2D Summary Expression ofthe NOV8 gene is ubiquitous among the samples in this panel, with highest expression in a kidney cancer sample (CT=27.1). Thus, the expression of this gene could be used to distinguish between this kidney cancer and the other samples in the panel.

Panel 4D Summary The NOV8 gene is widely expressed among the samples in this panel, with highest expression in endothelial (HUVEC) cells (CT=26.7). Significantly, expression ofthe NOV8 gene is higher in in activated B cells than in resting B cells. Small molecule drugs that inhibit the function ofthe NOV8 gene product may reduce the function of activated B cells and may reduce or eliminate the symptoms in patients with autoimmune and inflammatory diseases such as lupus erythematosus, Crohn's disease, ulcerative colitis, asthma, psoriasis, and rheumatoid arthritis.

Panel CNS_neurodegeneration_vl.O Summary The NOV8 gene is widely expressed among the samples in this panel, with highest expression in the temporal cortex of an Alzheimer's patient (CT=26.6). While the expression of this gene does not appear be specific to Alzheimer's disease, this panel confirms expression at the moderate/high level in the brain in an additional set of individuals (Voets et al., Muncl 8-1 promotes large dense-core vesicle docking. Neuron. 31(4):581-91, 2001).

NOV9: Immunoglobulin-like

Expression ofthe NOV9a gene (SCI 38673511_A) and the NOV9b variant (CG106625-02) was assessed using the primer-probe set Ag2176 described in Table 65. Results from RTQ-PCR runs are shown in Tables 66-70.

Table 65. Probe Name Ag2176

Table 66. Panel 1.3D

Table 67. Panel 2D

Table 68. Panel 4D

Table 69. Panel CNS 1

Panel 1.3D Summary Expression ofthe NOV9A gene is ubiquitous among the samples in this panel, with highest expression seen in the hippocampus (CT=25.1). The highly brain-preferential pattern of expression of this gene indicates a role in normal adult brain function. Please see Panel CNS_1 for further discussion of potential utility in the central nervous system.

Among tissues with metabolic function, the NOV9A gene has moderate expression in a variety of metabolic tissues including adrenal gland, adult and fetal heart, adult and fetal liver, adult and fetal skeletal muscle, pancreas and thyroid. Thus, this gene product may be a protein therapeutic for the treatment of diseases that affect these tissues, including obesity and diabetes.

Panel 2D Summary The NOV9A gene is widely expressed among the samples in this panel, with highest expression detected in normal kidney tissue adjacent to a kidney cancer (CT=28.7). In addition, other tissue samples derived from normal margin to kidney cancers appear to express this gene. Thus, the expression of this gene could be used to distinguish normal kidney tissue adjacent to malignant kidney. Moreover, therapeutic modulation ofthe expression or function of this gene or gene product, through the use of small molecule drugs, antibodies or protein therapeutics, might be of benefit in the treatment of kidney cancer.

Panel 4D Summary The results from two experiments with the same probe and primer set are in very good agreement, with highest expression in astrocytes (CTs=27-28).

Furthermore, expression ofthe NOV9A gene, which encodes a putative immunoglobulin domain containing protein, is higher in activated astrocytes when compared to resting astrocytes. Activated astrocytes play a role in the pathogenesis of multiple sclerosis (See reference 1). Thus, the NOV9A gene product could be used as a protein therapeutic for the reduction or elimination ofthe symptoms in patients suffering from multiple sclerosis.

Panel CNS_1 Summary This gene appears to be widely expressed across all regions ofthe brain and in all disease states represented in this panel. Apparent reduced expression in the primary motor strip (BA4 region) of an Alzheimer's brain suggests an association ofthe function ofthe NOV9A gene product with this neurodegenerative pathology. In addition, a less pronounced, but still evident, decreased expression ofthe NON9A gene in the primary motor strip (BA4) of Huntington's disease brains suggests a possible general role in neurodegeneration in this brain region. The combination of brain-preferential expression and dysregulation in neurodegenerative diseases seen in this panel indicates potential utility ofthe NOV9A gene product as a specific mediator of CNS disorders. Therefore, agents that modulate the action ofthe protein encoded by the NOV9A gene may have utility in the treatment of CNS disorders.

Panel CNS_neurodegeneration_V1.0 Summary The NOV9A gene is expressed across all regions ofthe brain, with highest expression detected in the occipital cortex of a control patient (CT=25.5). The gene does not appear to be differentially expressed in Alzheimer's disease. Please see Panel CNS_1 for potential utility in the central nervous system (Satoh and Kuroda Differing effects of IFN beta vs IFN gamma in MS: gene expression in cultured astrocytes. Neurology. 57:681-5, 2001).

NOVlOa: Type II Cytokeratin

Expression ofthe NOVlOa gene (GSAC055715.12_D) was assessed using the primer- probe set Agl858 described in Table 71. Results from RTQ-PCR runs are shown in Tables 72 and 73.

Table 71. Probe Name Agl858

Table 72. Panel 1.3D

Table 73. Panel 4D

Panel 1.3D Summary Expression ofthe NOV 10A gene is limited to a sample derived from a colon cancer cell line (CT=31.4) in this panel. Of note is the difference between SW480 cells and the genetically related metastatic variant, SW620, which showed much lower expression of this gene. Thus, the expression of this gene could be used to distinguish SW480 cells from the rest ofthe samples on the panel, and specifically the SW620 metastatic variant.

Panel 2.2 Summary Expression ofthe NOV 10A gene is low/undetectable (Ct values >35) in all samples in Panels 2.2. (Data not shown.)

Panel 4D Summary Expression ofthe NOV10A gene in panel 4D is limited to IL-13 stimulated lung fibroblasts (CT=33.5) and the mucoepidermoid cell line NCI-H292. The

NOV 10A gene, a basic cytokeratin homolog, may be useful as a small molecule target for the discovery of therapeutics to reduce or eliminate the symptoms of lung diseases such as asthma, emphysema, and chronic obstructive pulmonary disease. Furthermore, the NOV 10A gene product may be useful as a diagnostic marker for the cells in these diseases. NOVl 0b : Type II Cytokeratin Expression ofthe NOVlOb gene (GSAC055715_C) was assessed using the primer- probe set Agl857 described in Table 74. Results from RTQ-PCR runs are shown in Tables 75-

77.

Table 75. Panel 1.3D

Table 77. Panel 4D

Panel 1.3D Summary Expression ofthe NOVlOb gene is highest in the testis (CT=29.4), with low but significant expression also detected in the lung, lymph node and small intestine. Thus, the expression ofthe NOVlOb gene could be used to distinguish testis from the other samples in the panel.

Panel 2D Summary Expression ofthe NOVlOb gene is highest in a sample from normal breast tissue adjacent to a breast cancer (CT=32.9) with low but significant expression also detected in the breast cancer, bladder cancer, normal lung tissue, and a sample derived from normal tissue adjacent to a colon cancer. Thus, the expression ofthe NOVlOb gene could be used to distinguish these tissue samples from the rest ofthe samples in the panel.

Panel 4D Summary Expression ofthe NOVlOb gene is highest in resting Thl lymphocytes (CT=30.7), with significant expression also seen in resting Th2 cells. Lower levels of expression levels are detected in activated Thl and Th2 cells. The NOVlOb gene encodes a basic cytokeratin homolog and may be useful as a small molecule target for the discovery of therapeutics that can reduce or eliminate the symptoms of autoimmune and inflammatory diseases such as lupus erythematosus, Crohn's disease, ulcerative colitis, asthma, psoriasis, and rheumatoid arthritis. Furthermore, the NOVlOb gene product may be useful as a diagnostic marker for Thl and Th2 lymphocytes.

The NOVlOb gene and the NOVlOc gene (please see next entry) that encode these cytokeratin homologs are both found in the cytokeratin locus on chromosome 12. However, while all cytokeratins to date have been identified as epithelial cell markers, these novel gene products are expressed in a restricted subset of T lymphocytes, as seen from the expression profile in Panel 4D. CLUSTALW alignment of these two novel lymphocyte cytokeratins is shown in Table 10G and demonstrates similarity along the entire length ofthe proteins.

NOVlOc: Type II Cytokeratin

Expression ofthe NOVlOc gene (GSAC055715_B) was assessed using the primer- probe set Agl856 described in Table 78. Results from RTQ-PCR runs are shown in Tables 79- 81.

Table 78. Probe Name Agl856

Table 79. Panel 1.3D

Table 80. Panel 2D

Table 81. Panel 4D

(Endothelial) starved

Panel 1.3D Summary Highest expression ofthe NOVlOc gene is detected in the small intestine (CT=31.4). The expression of this gene is highest in a sample derived from small intestine. In addition, this gene has an expression pattern restricted to lymph node, bone marrow, spleen, thymus, trachea, adult lung and fetal lung tissue. Thus the expression of this gene could be used to distinguish these tissues from other tissues in the panel.

Panel 2D Summary Highest expression ofthe NOVlOc gene is seen in normal breast tissue adjacent to a breast cancer (CT=31.8). Low, but significant expression is also seen in normal lung tissue, as is seen in panel 1.3D and in some breast cancer samples. Thus, the expression of this gene could be used to distinguish these tissue samples from the rest ofthe samples in the panel. Moreover, therapeutic modulation ofthe expression of this gene or the function of its protein product, through the use of small molecule drugs, antibodies or protein therapeutics, might be of benefit for the treatment of breast cancer.

Panel 4D Summary Highest expression ofthe gene is seen in untreated CD4 cells (CT=28.8). Significant expression is also seen in resting Thl and Th2 lymphocytes, with lower levels of expression in activated Thl and Th2 cells. The NOVlOc gene encodes a cytokeratin homolog and may be useful as a small molecule target for the discovery of therapeutics that can reduce or eliminate the symptoms of autoimmune and inflammatory diseases such as lupus erythematosus, Crohn's disease, ulcerative colitis, asthma, psoriasis, and rheumatoid arthritis. Furthermore, the NOVlOc gene may be useful as a diagnostic marker for Thl and Th2 lymphocytes.

OTHER EMBODIMENTS

Although particular embodiments have been disclosed herein in detail, this has been done by way of example for purposes of illustration only, and is not intended to be limiting with respect to the scope ofthe appended claims, which follow. In particular, it is contemplated by the inventors that various substitutions, alterations, and modifications may be made to the invention without departing from the spirit and scope ofthe invention as defined by the claims. The choice of nucleic acid starting material, clone of interest, or library type is believed to be a matter of routine for a person of ordinary skill in the art with knowledge ofthe embodiments described herein. Other aspects, advantages, and modifications considered to be within the scope ofthe following claims.

Claims

WHAT IS CLAIMED IS:

1. An isolated polypeptide comprising an amino acid sequence selected from the group consisting of:

(a) a mature form of an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48;

(b) a variant of a mature form of an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48, wherein one or more amino acid residues in said variant differs from the amino acid sequence of said mature form, provided that said variant differs in no more than 15% ofthe amino acid residues from the amino acid sequence of said mature form;

(c) an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48; and

(d) a variant of an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48 wherein one or more amino acid residues in said variant differs from the amino acid sequence of said mature form, provided that said variant differs in no more than 15% of amino acid residues from said amino acid sequence.

2 The polypeptide of claim 1, wherein said polypeptide comprises the amino acid sequence of a naturally-occurring allelic variant of an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48.

3. The polypeptide of claim 2, wherein said allelic variant comprises an amino acid sequence that is the translation of a nucleic acid sequence differing by a single nucleotide from a nucleic acid sequence selected from the group consisting of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and/or 47.

4. The polypeptide of claim 1, wherein the amino acid sequence of said variant comprises a conservative amino acid substitution.

5. An isolated nucleic acid molecule comprising a nucleic acid sequence encoding a polypeptide comprising an amino acid sequence selected from the group consisting of:

(c) an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48;

(d) a variant of an amino acid sequence selected from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48, wherein one or more amino acid residues in said variant differs from the amino acid sequence of said mature form, provided that said variant differs in no more than 15% of amino acid residues from said amino acid sequence;

(e) a nucleic acid fragment encoding at least a portion of a polypeptide comprising an amino acid sequence chosen from the group consisting of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48, or a variant of said polypeptide, wherein one or more amino acid residues in said variant differs from the amino acid sequence of said mature form, provided that said variant differs in no more than 15% of amino acid residues from said amino acid sequence; and (f) a nucleic acid molecule comprising the complement of (a), (b), (c), (d) or (e).

6. The nucleic acid molecule of claim 5, wherein the nucleic acid molecule comprises the nucleotide sequence of a naturally-occurring allelic nucleic acid variant.

7. The nucleic acid molecule of claim 5, wherein the nucleic acid molecule encodes a polypeptide comprising the amino acid sequence of a naturally-occurring polypeptide variant.

8. The nucleic acid molecule of claim 5, wherein the nucleic acid molecule differs by a single nucleotide from a nucleic acid sequence selected from the group consisting of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and/or 47.

9. The nucleic acid molecule of claim 5, wherein said nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of

(a) a nucleotide sequence selected from the group consisting of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and/or 47;

(b) a nucleotide sequence differing by one or more nucleotides from a nucleotide sequence selected from the group consisting of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and/or 47, provided that no more than 20% of the nucleotides differ from said nucleotide sequence;

(c) a nucleic acid fragment of (a); and

(d) a nucleic acid fragment of (b).

10. The nucleic acid molecule of claim 5, wherein said nucleic acid molecule hybridizes under stringent conditions to a nucleotide sequence chosen from the group consisting of SEQ ID NOS:l, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45 and/or 47, or a complement of said nucleotide sequence.

11. The nucleic acid molecule of claim 5, wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of

(a) a first nucleotide sequence comprising a coding sequence differing by one or more nucleotide sequences from a coding sequence encoding said amino acid sequence, provided that no more than 20% ofthe nucleotides in the coding sequence in said first nucleotide sequence differ from said coding sequence;

(b) an isolated second polynucleotide that is a complement ofthe first polynucleotide; and

(c) a nucleic acid fragment of (a) or (b).

12. A vector comprising the nucleic acid molecule of claim 11.

13. The vector of claim 12, further comprising a promoter operably-linked to said nucleic acid molecule.

14. A cell comprising the vector of claim 12.

15. An antibody that immunospecifically-binds to the polypeptide of claim 1.

16. The antibody of claim 15 , wherein said antibody is a monoclonal antibody.

17. The antibody of claim 15, wherein the antibody is a humanized antibody.

18. A method for determining the presence or amount ofthe polypeptide of claim 1 in a sample, the method comprising:

(a) providing the sample;

(b) contacting the sample with an antibody that binds immunospecifically to the polypeptide; and

(c) determining the presence or amount of antibody bound to said polypeptide, thereby determining the presence or amount of polypeptide in said sample.

19. A method for determining the presence or amount ofthe nucleic acid molecule of claim 5 in a sample, the method comprising:

(a) providing the sample;

(b) contacting the sample with a probe that binds to said nucleic acid molecule; and

(c) determining the presence or amount ofthe probe bound to said nucleic acid molecule, thereby determining the presence or amount of the nucleic acid molecule in said sample.

20. A method of identifying an agent that binds to a polypeptide of claim 1 , the method comprising:

(a) contacting said polypeptide with said agent; and

(b) determining whether said agent binds to said polypeptide.

21. A method for identifying an agent that modulates the expression or activity of the polypeptide of claim 1, the method comprising:

(a) providing a cell expressing said polypeptide;

(b) contacting the cell with said agent; and

(c) determining whether the agent modulates expression or activity of said polypeptide, whereby an alteration in expression or activity of said peptide indicates said agent modulates expression or activity of said polypeptide.

22. A method for modulating the activity ofthe polypeptide of claim 1, the method comprising contacting a cell sample expressing the polypeptide of said claim with a compound that binds to said polypeptide in an amount sufficient to modulate the activity ofthe polypeptide.

23. A method of treating or preventing a NOVX-associated disorder, said method comprising administering to a subject in which such treatment or prevention is desired the polypeptide of claim 1 in an amount sufficient to treat or prevent said NOVX- associated disorder in said subject.

24. The method of claim 23, wherein said subject is a human.

25. A method of treating or preventing a NOVX-associated disorder, said method comprising administering to a subject in which such treatment or prevention is desired the nucleic acid of claim 5 in an amount sufficient to treat or prevent said NOVX- associated disorder in said subject.

26. The method of claim 25, wherein said subject is a human.

27. A method of treating or preventing a NOVX-associated disorder, said method comprising administering to a subject in which such treatment or prevention is desired the antibody of claim 15 in an amount sufficient to treat or prevent said NOVX- associated disorder in said subject.

28. The method of claim 27, wherein the subject is a human.

29. A pharmaceutical composition comprising the polypeptide of claim 1 and a pharmaceutically-acceptable carrier.

30. A pharmaceutical composition comprising the nucleic acid molecule of claim 5 and a pharmaceutically-acceptable carrier.

31. A pharmaceutical composition comprising the antibody of claim 15 and a pharmaceutically-acceptable carrier.

32. A kit comprising in one or more containers, the pharmaceutical composition of claim 29.

33. A kit comprising in one or more containers, the pharmaceutical composition of claim 30.

34. A kit comprising in one or more containers, the pharmaceutical composition of claim 31.

35. The use of a therapeutic in the manufacture of a medicament for treating a syndrome associated with a human disease, the disease selected from a NOVX-associated disorder, wherein said therapeutic is selected from the group consisting of a NOVX polypeptide, a NOVX nucleic acid, and a NOVX antibody.

36. A method for screening for a modulator of activity or of latency or predisposition to a NOVX-associated disorder, said method comprising:

(a) administering a test compound to a test animal at increased risk for a NOVX-associated disorder, wherein said test animal recombinantly expresses the polypeptide of claim 1 ;

(b) measuring the activity of said polypeptide in said test animal after administering the compound of step (a);

(c) comparing the activity of said protein in said test animal with the activity of said polypeptide in a control animal not administered said polypeptide, wherein a change in the activity of said polypeptide in said test animal relative to said control animal indicates the test compound is a modulator of latency of or predisposition to a NOVX-associated disorder.

37. The method of claim 36, wherein said test animal is a recombinant test animal that expresses a test protein transgene or expresses said transgene under the control of a promoter at an increased level relative to a wild-type test animal, and wherein said promoter is not the native gene promoter of said transgene.

38. A method for determining the presence of or predisposition to a disease associated with altered levels ofthe polypeptide of claim 1 in a first mammalian subject, the method comprising:

(a) measuring the level of expression of the polypeptide in a sample from the first mammalian subject; and

(b) comparing the amount of said polypeptide in the sample of step (a) to the amount ofthe polypeptide present in a control sample from a second mammalian subject known not to have, or not to be predisposed to, said disease, wherein an alteration in the expression level ofthe polypeptide in the first subject as compared to the control sample indicates the presence of or predisposition to said disease.

39. A method for determining the presence of or predisposition to a disease associated with altered levels of the nucleic acid molecule of claim 5 in a first mammalian subject, the method comprising:

(a) measuring the amount ofthe nucleic acid in a sample from the first mammalian subject; and

(b) comparing the amount of said nucleic acid in the sample of step (a) to the amount ofthe nucleic acid present in a control sample from a second mammalian subject known not to have or not be predisposed to, the disease; wherein an alteration in the level ofthe nucleic acid in the first subject as compared to the control sample indicates the presence of or predisposition to the disease.

40. A method of treating a pathological state in a mammal, the method comprising administering to the mammal a polypeptide in an amount that is sufficient to alleviate the pathological state, wherein the polypeptide is a polypeptide having an amino acid sequence at least 95% identical to a polypeptide comprising an amino acid sequence of at least one of SEQ ID NOS:2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, 40, 42, 44, 46 and/or 48, or a biologically active fragment thereof.

1. A method of treating a pathological state in a mammal, the method comprising administering to the mammal the antibody of claim 15 in an amount sufficient to alleviate the pathological state.