title: Characterization of the Ah receptor-associated protein, ARA9.
abstract: The unliganded aryl hydrocarbon receptor (AHR) is found in a complex with other proteins including the 90-kDa heat shock protein (Hsp90) and a 37-kDa protein we refer to as ARA9. We found that the three tetratricopeptide repeats found in the COOH terminus of ARA9 are necessary and sufficient for interaction with the AHR complex. Conversely, the AHR's "repressor"/Hsp90 binding domain is required for interaction with ARA9. Because ARA9 closely resembles the 52-kDa FK506-binding protein (FKBP52), found in the unliganded glucocorticoid receptor (GR) complex, we compared the binding specificities of ARA9 and FKBP52 for AHR and GR. In co-immunoprecipitation experiments, ARA9 specifically associated with AHR-Hsp90 complex but not with GR-Hsp90 complexes. In addition, ARA9 showed a greater capacity than FKBP52 to associate with AHR-Hsp90 complexes. The biological importance of this interaction was suggested by the observation that in a yeast expression system ARA9 expression enhanced the response of AHR to the agonist beta-napthoflavone, decreasing the EC50 by greater than 5-fold and increasing the maximal response 2.5-fold. In contrast, co-expression of FKBP52 had no effect on AHR signaling. In addition, although ARA9 contains a domain similar to that found in other FK506-binding proteins, ARA9 binding to 3H-FK506 could not be detected. Finally, we have characterized the developmental and expression pattern of ARA9 in the developing mouse embryo and mapped the ARA9 locus to human chromosome 11q13.3.
authors.affiliation: McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison, Wisconsin 53706, USA.
authors.forename: L A
authors.initials: LA
authors.lastname: Carver
authors.forename: J J
authors.initials: JJ
authors.lastname: LaPres
authors.forename: S
authors.initials: S
authors.lastname: Jain
authors.forename: E E
authors.initials: EE
authors.lastname: Dunham
authors.forename: C A
authors.initials: CA
authors.lastname: Bradfield
mesh_terms: D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002874:Chromosome Mapping; D017931:DNA Primers; D018507:Gene Expression Regulation, Developmental; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D051379:Mice; D011485:Protein Binding; D011506:Proteins; D018336:Receptors, Aryl Hydrocarbon / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016559:Tacrolimus / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D017931:DNA Primers; D018841:HSP90 Heat-Shock Proteins; D047908:Intracellular Signaling Peptides and Proteins; D011506:Proteins; D018336:Receptors, Aryl Hydrocarbon; C103265:aryl hydrocarbon receptor-interacting protein; D016559:Tacrolimus