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 . (canceled).  
     
     
         2 . (Canceled).  
     
     
         3 . (Canceled).  
     
     
         4 . (Canceled).  
     
     
         5 . (Canceled).  
     
     
         6 . (Canceled).  
     
     
         7 . (Canceled).  
     
     
         8 . (Canceled).  
     
     
         9 . (Canceled).  
     
     
         10 . (Canceled).  
     
     
         11 . (Canceled).  
     
     
         12 . (Canceled).  
     
     
         13 . (Canceled).  
     
     
         14 . (Canceled).  
     
     
         15 . (Canceled).  
     
     
         16 . (Canceled).  
     
     
         17 . (Canceled).  
     
     
         18 . An isolated estrogen recepter-β comprising amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID No. 2.  
     
     
         19 . (Canceled).  
     
     
         20 . (Canceled).  
     
     
         21 . (Canceled).  
     
     
         22 . (Canceled).  
     
     
         23 . An antibody that specifically recognizes hERβ.  
     
     
         24 . A method for identifying hERβ-interactive compounds, said method comprising: 
 (a) contacting the polypeptide of  claim 18 , which polypeptide encodes hERβ, with a labeled ligand in the presence of test compounds, to form test reactions, and in the absence of test compounds, to form control reactions;  
 (b) incubating said test and control reactions under appropriate conditions to achieve equilibrium binding of said labeled ligand to hERβ;  
 (c) determining the level of binding of said labeled ligand to hERβ in said test and control cultures; and  
 (d) identifying as a hERβ-interactive compound any compound that reduces the binding of said labeled ligand to hERβ.  
 
     
     
         25 . A method as defined in  claim 24 , wherein said ligand is 17-β estradiol.  
     
     
         26 . A method as defined in  claim 24 , wherein said hERβ-interactive compound is an agonist.  
     
     
         27 . A method as defined in  claim 24 , wherein said hERβ-interactive compound is an antagonist.  
     
     
         28 . (Canceled).  
     
     
         29 . (Canceled).  
     
     
         30 . (Canceled).  
     
     
         31 . The polypeptide of  claim 18 , wherein the polypeptide is modified with a label capable of providing a detectable signal.  
     
     
         32 . The polypeptide of  claim 31 , wherein the label is a radioisotope.  
     
     
         33 . The polypeptide of  claim 31 , wherein the label is a fluorescent compound.  
     
     
         34 . (Canceled).  
     
     
         35 . (Canceled).  
     
     
         36 . The polypeptide of  claim 18 , wherein the polypeptide is produced in intact cells.  
     
     
         37 . The polypeptide of  claim 18 , wherein the polypeptide is produced in cell-free translation systems.  
     
     
         38 . (Canceled).  
     
     
         39 . (Canceled).  
     
     
         40 . The polypeptide of  claim 18 , wherein the polypeptide is chemically synthesized.  
     
     
         41 . The polypeptide of  claim 18 , wherein the polypeptide is produced in a recombinant system.  
     
     
         42 . A purified polypeptide comprising amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID NO:2, wherein when this polypeptide forms the N-terminus of human estrogen receptor β, the estrogen receptor β stimulates estrogen response element (ERE) activity to a greater extent than the truncated estrogen receptor lacking this N-terminal polypeptide sequence.  
     
     
         43 . A purified polypeptide comprising amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID NO:2, wherein when this polypeptide forms the N-terminus of human estrogen receptor β, the estrogen receptor β attenuates NF-kB transcription activation while the truncated estrogen receptor lacking this N-terminal polypeptide sequence does not.  
     
     
         44 . A purified polypeptide comprising amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID NO:2, wherein when this polypeptide forms the N-terminus of human estrogen receptor β, the estrogen receptor β is 2 to 3 times more active than the truncated estrogen receptor lacking this N-terminal polypeptide sequence in activating the ERE-reporter gene in the presence of estradiol.  
     
     
         45 . An isolated estrogen receptor-β comprising an N-terminus having amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID NO:2, wherein the estrogen receptor β stimulates ERE activity to a greater extent than the truncated estrogen receptor lacking this N-terminal polypeptide sequence.  
     
     
         46 . An isolated estrogen receptor-β comprising an N-terminus having amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID NO:2, wherein the estrogen receptor β attenuates NF-kB transcription activation while the truncated estrogen receptor lacking this N-terminal polypeptide sequence does not.  
     
     
         47 . An isolated estrogen receptor-β comprising an N-terminus having amino acids 1-45 of the sequence depicted in FIG. 4 SEQ ID NO:2, wherein the estrogen receptor β is 2 to 3 times more active than the truncated estrogen receptor lacking this N-terminal polypeptide sequence in activating the ERE-reporter gene in the presence of estradiol.