Human MAD proteins and uses thereof

Abstract

Human MADr3 or MADr4 polypeptides and DNA (RNA) encoding such MADr3 or MADr4 and a procedure for producing such polypeptides by recombinant techniques is disclosed. Also disclosed are methods for utilizing such MADr3 or MADr4, or compounds which inhibit or stimulate MADr3 or MADr4 for stimulating wound healing, and treating cancers, among others, are also disclosed. Agonist and antagonists of these MAD proteins and methods of their use are also disclosed. Also disclosed are diagnostic assays for detecting diseases related to mutations in the nucleic acid sequences and altered concentrations of the polypeptides. Also disclosed are diagnostic assays for detecting mutations in the polynucleotides encoding the MADr3 or MADr4 and for detecting altered levels of the polypeptide in a host.

What is claimed is:  
     
         1 . An isolated polynucleotide comprising a member selected from the group consisting of: 
 (a) a polynucleotide encoding the polypeptide as set forth in SEQ ID NO: 2;    (b) a polynucleotide having at least 70% identity to a polynucleotide encoding a polypeptide comprising amino acids of SEQ ID NO: 2;    (c) a polynucleotide which by virtue of the redundancy of the genetic code, encodes the same amino acids of SEQ ID NO: 2;    (d) a polynucleotide encoding the polypeptide comprising amino acids 1 to 425 of FIGS.  1 A- 1 D;    (e) a polynucleotide which is complementary to the polynucleotide of (a) through (d), and    (f) a polynucleotide comprising at least 15 contiguous bases of the polynucleotide of (a) through (e).    
     
     
         2 . The polynucleotide of  claim 1  wherein the polynucleotide is DNA or RNA.  
     
     
         3 . The polynucleotide of  claim 2  comprising nucleotides set forth in SEQ ID NO: 1.  
     
     
         4 . The polynucleotide of  claim 2  comprising nucleotides 1-2333 set forth in SEQ ID NO: 1.  
     
     
         5 . The polynucleotide of  claim 2  which encodes a polypeptide comprising the amino acids of SEQ ID NO: 2.  
     
     
         6 . An isolated polynucleotide comprising a member selected from the group consisting of: 
 (a) a polynucleotide as set forth in SEQ ID NO: 3;    (b) a polynucleotide as set forth in SEQ ID NO: 4;    (c) a polynucleotide having at least 70% identity to a polynucleotide of (a) or (b);    (d) a polynucleotide which by virtue of the redundancy of the genetic code, encodes the same amino acids as may be encoded by SEQ ID NO: 3 or 4;    (e) a polynucleotide which is complementary to the polynucleotide of (a) through (d), and    (f) a polynucleotide comprising at least 15 contiguous bases of the polynucleotide of (a) through (e).    
     
     
         7 . The polynucleotide of  claim 7  wherein the polynucleotide is DNA or RNA.  
     
     
         8 . The polynucleotide of  claim 7  comprising nucleotides set forth in SEQ ID NO: 3.  
     
     
         9 . The polynucleotide of  claim 7  comprising nucleotides set forth in SEQ ID NO: 4.  
     
     
         10 . An isolated polynucleotide comprising a member selected from the group consisting of: 
 (a) a polynucleotide which encodes a mature polypeptide having the amino acid sequence expressed by the DNA contained in ATCC Deposit No. 98223;    (b) a polynucleotide having at least a 70% identity to a polynucleotide encoding the same mature polypeptide of (a);    (c) a polynucleotide which by virtue of the redundancy of the genetic code encodes the same mature polypeptide expressed by the human cDNA contained in ATCC Deposit No. 98223    (d) a polynucleotide complementary to the polynucleotide of (a) through (c); and    (e) a polynucleotide comprising at least 15 contiguous bases of the polynucleotide of (a) through (d).    
     
     
         11 . A vector comprising the DNA of  claim 2 .  
     
     
         12 . A vector comprising the DNA of  claim 7 .  
     
     
         13 . A host cell comprising the vector of  claim 11 .  
     
     
         14 . A host cell comprising the vector of  claim 12 .  
     
     
         15 . A process for producing a polypeptide comprising expressing from the host cell of  claim 13  a polypeptide encoded by said DNA.  
     
     
         16 . A process for producing a cell which expresses a polypeptide comprising transforming or transfecting the cell with the vector of  claim 11  such that the cell expresses the polypeptide encoded by the human cDNA contained in the vector.  
     
     
         17 . A polypeptide comprising an amino acid sequence which is at least 70% identical to the amino acid sequence of SEQ ID NO: 2.  
     
     
         18 . A polypeptide comprising an amino acid sequence as set forth in SEQ ID NO: 2.  
     
     
         19 . A compound which inhibits activation of the polypeptide of  claim 17 .  
     
     
         20 . An antibody against the polypeptide of  claim 17 .  
     
     
         21 . A compound which activates the polypeptide of  claim 17 .  
     
     
         22 . A method for the treatment of a patient having need of MADr3 comprising administering to the patient a therapeutically effective amount of the polypeptide of  claim 17 .  
     
     
         23 . The method of  claim 22  wherein said therapeutically effective amount of the polypeptide is administered by providing to the patient DNA encoding said polypeptide and expressing said polypeptide in vivo.  
     
     
         24 . A method for the treatment of a patient having need to inhibit MADr3 or MADr4 polypeptide comprising administering to the patient a therapeutically effective amount of the compound of  claim 19 .  
     
     
         25 . A process for diagnosing a disease or a susceptibility to a disease related to expression of the polypeptide of  claim 17  comprising determining a mutation in the nucleic acid sequence encoding said polypeptide.  
     
     
         26 . A diagnostic process comprising analyzing for the presence of the polypeptide of  claim 17  in a sample derived from a host.  
     
     
         27 . A method for identifying compounds which bind to and activate or inhibit a ligand of the polypeptide of  claim 12  comprising: 
 contacting a cell expressing on the surface thereof a ligand for the polypeptide, said ligand being associated with a second component capable of providing a detectable signal in response to the binding of a compound to said ligand, with a compound to be screened under conditions to permit binding to the ligand; and  
 determining whether the compound binds to and activates or inhibits the ligand by detecting the presence or absence of a signal generated from the interaction of the compound with the ligand.  
 
     
     
         28 . A process for identifying compounds active as agonists to the polypeptide of  claim 17  comprising: contacting a reaction mixture containing a cell type which expresses a MADr3 ligand and a compound to be screened; and determining if the compound generates a signal from said MADr3 or MADr4 to identify if the compound is an effective agonist.  
     
     
         29 . A process for identifying compounds active as antagonists which inhibit phosphorylation of the polypeptide of  claim 17 , inhibit interaction with other proteins, inhibit the translocation of MADr3 to the nucleus and inhibit the interaction of MADr3 with nucleic acid sequences comprising: 
 contacting a reaction mixture containing a cell type which expresses the MADr3 ligand and a compound to be screened; and detecting the absence of a signal generated from said ligand after binding of said compound to identify if the compound is an effective antagonist.