Novel card proteins involved in cell death regulation

Abstract

The present invention provides NB-ARC and CARD-containing proteins (NACs), nucleic acid molecules encoding NACs and antibodies specific for at least one NAC. The invention further provides chimeric NAC proteins. The invention also provides screening assays for identifying an agent that can effectively alter the association of a NAC with a NAC-associated protein. The invention further provides methods of modulating apoptosis in a cell by introducing into the cell a nucleic acid molecule encoding a NAC or an antisense nucleotide sequence. The invention also provides a method of using a reagent that can specifically bind to a NAC to diagnose a pathology that is characterized by an increased or decreased level of apoptosis in a cell.

1 . An isolated anti-NAC antibody having specific reactivity with a NAC, said NAC comprising a NB-ARC domain, a CARD domain and a TIM-Barrel-like domain. 
     
     
         2 . Antibody according to  claim 1 , wherein said antibody is a monoclonal antibody. 
     
     
         3 . A cell line producing the monoclonal antibody of  claim 2 . 
     
     
         4 . An antibody according to  claim 1 , wherein said antibody is a polyclonal antibody. 
     
     
         5 . A therapeutic composition comprising a compound selected from a NAC, or functional fragment thereof, a NAC modulating agent, or an anti-NAC antibody; and a pharmaceutically acceptable carrier, said NAC modulating agent identified by a method comprising:
 a) contacting said NAC and NAP proteins, under conditions that allow said NAC and NAP proteins to associate, with an agent suspected of being able to alter the association of said NAC and NAP proteins; and   b) detecting the altered association of said NAC and NAP proteins, wherein said altered association identifies an effective agent.   
     
     
         6 . A method of treating a pathology characterized by abnormal cell proliferation or abnormal inflammation, said method comprising administering an effective amount of the composition according to  claim 5 . 
     
     
         7 . A method of diagnosing a pathology characterized by an increased or decreased level of a NAC in a subject, comprising the steps of:
 a) obtaining a test sample from the subject;   b) contacting said test sample with an agent that can bind said NAC under suitable conditions, which allow specific binding of said agent to said NAC; and   c) comparing the amount of said specific binding in said test sample with the amount of specific binding in a control sample, wherein an increased or decreased amount of said specific binding in said test sample as compared to said control sample is diagnostic of a pathology.   
     
     
         8 . The method of  claim 7 , wherein said agent is an anti-NAC antibody or a NAC-associated-protein (NAP). 
     
     
         9 . A method of modulating transcription comprising contacting a cell with a compound selected from the group consisting of: a NAC protein or functional fragment thereof, an agent, and an anti-NAC antibody, said agent identified by a method comprising steps (a) and (b) of  claim 5 . 
     
     
         10 . A method of diagnosing cancer or monitoring cancer therapy comprising contacting a test sample from a patient with the antibody of  claim 1 . 
     
     
         11 . A method of assessing prognosis of patients with cancer comprising contacting a test sample from a patient with the antibody of  claim 1 . 
     
     
         12 . An isolated protein selected from a protein comprising an NB-ARC domain, a CARD domain and a TIM-Barrel-like domain and a protein comprising an NB-ARC and CARD containing protein (NAC), wherein the amino acid sequence of said protein comprises substantially the same sequence as any of SEQ ID NOs: 2, 4 or 6. 
     
     
         13 . The protein of  claim 12 , further comprising a LRR domain. 
     
     
         14 . A NAC according to  claim 12  comprising the same amino acid sequence as set forth in any of SEQ ID NOs: 2, 4 or 6. 
     
     
         15 . A NAC according to  claim 12 , wherein said protein is encoded by a nucleotide sequence comprising substantially the same nucleotide sequence as set forth in SEQ ID NOs: 1, 3 or 5. 
     
     
         16 . A NAC according to  claim 12 , wherein said protein is encoded by a nucleotide sequence comprising the same sequence as set forth in SEQ ID NOs: 1, 3 or 5.