Abstract
Aryl hydrocarbon receptor (AhR) is a transcription factor that is activated by the binding of xenobiotic and endogenous ligands. AhR interacts with heat shock protein (Hsp) 90 complexes and can be used as a functional substrate to detect chaperone-dependent processes. Yeast Hsp90 (hsp82) mutants that variably affected AhR signaling were identified using reporter gene assays. Some mutated alleles resided in the p23/adenosine triphosphate (ATP)-binding pocket of Hsp90, so the relationship between the cochaperone Sba1 (yeast p23) and adenosine triphosphatase (ATPase) activity was investigated. Deletion of the p23 gene in the hsp82G170D mutant background had a greater effect on AhR signaling than the individual mutations, suggesting that these 2 mutations have separate actions on AhR signaling. In contrast, p23 overexpression suppressed temperature sensitivity and AhR signaling defects in the hsp82G170D mutant strain, suggesting that there is a relationship between these 2 proteins. The mutated hsp82G170D protein lacked detectable ATPase activity and p23 binding in vitro, which may relate to the weakened AhR signaling observed in mutant cells. Sba1 (p23) suppressed Hsp82 ATPase activity in vitro. These studies implicate the p23 protein and the G170 region of Hsp90 as being important, but not essential, for AhR signaling. Our results are consistent with a model in which p23 inhibits Hsp90 ATPase activity, thereby stabilizing ATP-Hsp90-client protein complexes.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Triphosphatases / genetics
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Adenosine Triphosphatases / metabolism
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Adenosine Triphosphate / analogs & derivatives*
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Adenosine Triphosphate / pharmacology
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Benzoquinones
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Binding Sites / genetics
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Blotting, Western
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Escherichia coli / genetics
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Gene Deletion
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Gene Expression Regulation, Fungal / drug effects
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Genes, Reporter / genetics
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Genotype
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HSP90 Heat-Shock Proteins / genetics
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HSP90 Heat-Shock Proteins / metabolism
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HSP90 Heat-Shock Proteins / physiology*
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Lactams, Macrocyclic
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Molecular Chaperones / genetics
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Molecular Chaperones / metabolism
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Molecular Chaperones / physiology*
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Plasmids / genetics
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Point Mutation / genetics
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Polymerase Chain Reaction
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Protein Binding / drug effects
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Quinones / pharmacology
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Receptors, Aryl Hydrocarbon / physiology*
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Recombinant Proteins / genetics
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Recombinant Proteins / metabolism
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Saccharomyces cerevisiae / drug effects
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Saccharomyces cerevisiae / genetics
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Saccharomyces cerevisiae / physiology*
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
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Saccharomyces cerevisiae Proteins / physiology*
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Signal Transduction / physiology
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Temperature
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Transformation, Genetic
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beta-Naphthoflavone / pharmacology
Substances
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Benzoquinones
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HSP90 Heat-Shock Proteins
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Lactams, Macrocyclic
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Molecular Chaperones
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Quinones
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Receptors, Aryl Hydrocarbon
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Recombinant Proteins
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SBA1 protein, S cerevisiae
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Saccharomyces cerevisiae Proteins
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adenosine 5'-O-(3-thiotriphosphate)
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beta-Naphthoflavone
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Adenosine Triphosphate
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Adenosine Triphosphatases
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geldanamycin