Ah receptor cDNA and genetically engineered cells for detecting agonists to the Ah receptor

Abstract

Murine and human Ah receptor cDNAs are provided. These molecules can be used to generate large quantities of Ah-receptor protein. The Ah receptor can be inserted into cell systems such as yeast or mammalian, expressed, and used in assays to detect agonists to the Ah receptor. The Ah receptor inserted into such systems can be either the full Ah receptor, the receptor containing deletions at its amino and carboxyl ends, or a chimeric receptor. The chimeric Ah receptor has its finding and dimerization domains replaced with an analogous region from another binding protein.

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         21 . An isolated and purified polypeptide comprising a DNA binding domain of an Ah receptor protein wherein said DNA binding domain comprises the amino acid sequence of 
 [RKRRKPVQKTVKPIPAEGIKSNPSKRHRDRLNTELDRLASLLPFPQDVINKLDKLSV LRLSVSYLRAKSFFD] SEQ ID NO: 37 and alterations, substitutions and deletions thereof.    
     
     
         22 . An isolated and purified polypeptide comprising a DNA binding domain of an Ah receptor protein wherein said DNA binding domain comprises the amino acid sequence of 
 [RKRRKPVQKTVKPIPAEGIKSNPSKRHRDRLNTELDRLASLLPFPQDVINKLDKLSV LRLSVSYLRAKSFFD] SEQ ID NO: 38 and alterations, substitutions and deletions thereof.    
     
     
         23 . A chimeric Ah receptor protein wherein the native DNA binding domain of said chimeric protein has been replaced with a non-native DNA binding domain, wherein said DNA binding domain is a fusion protein capable of binding a DNA sequence.  
     
     
         24 . The chimeric Ah receptor protein of  claim 23  wherein the non-native binding domain is replaced with a DNA binding domain from a LexA protein.  
     
     
         25 . The chimeric Ah receptor protein of  claim 23  wherein the non-native DNA binding domain is replaced with a DNA binding domain from a Ga14 protein.  
     
     
         26 . An isolated and purified polypeptide comprising a DNA binding domain of an Ah receptor protein wherein said DNA binding domain comprises the amino acid sequence of [EFQRQLHWALNPSQCTESGQGIEEATGLPQTVVCYNPDQIPPENSPLMERCFICR LRCLLDNSSGFLAMNFQGKLKYLHGQKKKGKDGSILPPQLALFAIATPLQPPSILEIR TKNFIFRTKHKLDFTPIGCDAKGRIVLGYTEAELCTRGSGYOFIHMDMLYCAESHIR MIKTGESGMIVFRLLTKNNRWTWVQSNARLLYKNGRP] SEQ ID NO:39 and alterations, substitutions, additions and deletions thereof.  
     
     
         27 . An isolated and purified polypeptide comprising a ligand binding domain of an Ah receptor protein, wherein said ligand binding domain comprises the amino acid sequence of [FMERCFRCRLRCLLDNSSGFLAMNFQRLKYLHGQNKKGKDGALLPPQLALFAIAT PLQPPSILEIRTKNFIFRTKHKLDFTPIGCDAKGQLILGYTEVELCTRGSGYOFIHAADI LHCAESHIRMIKTGESGMTVFRLLAKHSRWRWVQSNARLIYRNGRP] SEQ ID NO:  40  and alterations, substitutions, additions and deletions thereof.