Novel human estrogen receptor beta

Abstract

The present invention provides isolated nucleic acids encoding full-length human estrogen receptor-β (hERβ), which comprises 530 amino acids. The invention also provides isolated hERβ polypeptides and hERβ-reactive antibodies, including those that specifically recognize amino acids 1-45 of hERβ. The invention also encompasses methods for identifying hERβ-interactive compounds, including agonists, antagonists, and co-activators.

1 - 30 . (canceled)  
     
     
         31 . An isolated nucleic acid encoding a polypeptide, wherein said polypeptide comprises the amino acid sequence set forth in  FIG. 4  (SEQ ID NO:2).  
     
     
         32 . The isolated nucleic acid of  claim 31 , wherein said nucleic acid comprises the nucleotide sequence set forth in  FIG. 3  (SEQ ID NO:1).  
     
     
         33 . The isolated nucleic acid of  claim 31 , wherein said nucleic acid is DNA.  
     
     
         34 . The isolated nucleic acid of  claim 31 , wherein said nucleic acid is RNA.  
     
     
         35 . A recombinant vector comprising a nucleic acid according to  claim 31 .  
     
     
         36 . The recombinant vector of  claim 35 , wherein the nucleic acid is operably linked to a transcription regulatory element.  
     
     
         37 . A host cell comprising a recombinant vector according to  claim 35 .  
     
     
         38 . A host cell according to  claim 37 , wherein the cell is a bacterial cell, a fungal cell, a plant cell, an insect cell or a vertebrate cell.  
     
     
         39 . A host cell according to  claim 37 , wherein said cell expresses a polypeptide encoded by the recombinant vector.  
     
     
         40 . A method for producing a polypeptide, which method comprises incubating a cell according to  claim 37  under conditions that permit expression of a heterologous polypeptide encoded by the recombinant vector.  
     
     
         41 . The method of  claim 40  further comprising steps of: 
 (a) harvesting the incubated cells to produce (i) a cell fraction, and (ii) a medium fraction; and    (b) recovering the polypeptide from the cell fraction, the medium fraction or both.    
     
     
         42 . A method for identifying compounds that interact with hERβ polypeptide, wherein the HERβ polypeptide comprises the amino acid sequence set forth in  FIG. 4  (SEQ ID NO:2), and wherein said method comprises steps of: 
 (a) bringing a host cell into contact with one or more test compounds, said host cell being transformed with (i) an isolated nucleic acid encoding the hERβ polypeptide, and (ii) a reporter gene functionally linked to an estrogen-responsive promoter element; and    (b) monitoring expression of the reporter gene,    wherein a change in expression of the reporter gene identies a compound that interacts with the hERβ polypeptide.    
     
     
         43 . The method of  claim 42 , further comprising a step of introducing into the host cell (i) the nucleic acid encoding hERβ polypeptide, and (ii) the reporter gene functionally linked to the estrogen-responsive reporter element.  
     
     
         44 . The method of  claim 42 , wherein the nucleic acid encoding hERβ polypeptide comprises the nucleotide sequence set forth in  FIG. 3  (SEQ ID NO:1).  
     
     
         45 . The method of  claim 42 , wherein the estrogen-responsive promoter element is an estrogen response element.  
     
     
         46 . The method of  claim 42  wherein the host cell is a HepG2 cell.  
     
     
         47 . The method of  claim 42 , wherein the host cell is a HAECT-1 cell.  
     
     
         48 . The method of  claim 42 , wherein the host cell is a COS cell, a HeLa cell, a CHO cell, a HUVEC cell, or a yeast cell.  
     
     
         49 . The method of  claim 48 , wherein the host cell is a yeast cell.  
     
     
         50 . The method of  claim 42 , wherein the reporter gene is a luciferase gene, a chloramphenicol acetyl transferase (CAT) gene, or a green fluorescence protein gene.  
     
     
         51 . The method of  claim 50 , wherein the reporter gene is a luciferase gene.  
     
     
         52 . The method of  claim 42 , wherein the reporter gene is a β-galactosidase gene.  
     
     
         53 . A method for identifying compounds that interact with hERβ polypeptide, wherein the HERβ polypeptide comprises the amino acid sequence set forth in  FIG. 4  (SEQ ID NO:2), and wherein said method comprises steps of: 
 (a) introducing into a host cell (i) an isolated nucleic acid encoding the hERβ polypeptide, and (ii) a reporter gene functionally linked to an estrogen responsive promoter element;    (b) brining the host cell into contact with one or more test compounds; and    (c) monitoring expression of the reporter gene,    wherein a change in expression of the reporter gene identies a compound that interacts with the hERβ polypeptide.