In Saccharomyces cerevisiae, the heat shock transcription factor (HSF) is thought to be a homotypic trimer that is bound to the promoters of heat shock protein (HSP) genes at both normal and heat shock temperatures. Exposure to heat shock greatly and rapidly induces HSF transcriptional activity without further increasing DNA-binding affinity. It is believed that HSF is under negative regulation at normal growth temperatures, but the detailed mechanism by which HSF is activated is still not clear. We report the analysis of mutations in a conserved arginine (residue 274) at the C-terminal end of the DNA-binding domain (DBD). Two mutations significantly increase both basal activity of HSF at normal temperatures and induced activity on heat shock. We demonstrate by coimmunoprecipitation experiments that the mutations reduce the association between the DNA-binding domain/oligomerization domain and the transcription activation domains. Our studies suggest that the DNA-binding domain of HSF can interact with activation domains directly, and this interaction is important for the repression of HSF activity under normal growth conditions. Destabilizing this interaction by heat or by mutations results in HSF transcriptional activation. We propose that Arg-274 is critical for intramolecular repression of HSF activity in normally growing cells.
|Evidence ID||Analyze ID||Interactor||Interactor Systematic Name||Interactor||Interactor Systematic Name||Type||Assay||Annotation||Action||Modification||Phenotype||Source||Reference||Note|
|Evidence ID||Analyze ID||Gene||Gene Systematic Name||Gene Ontology Term||Gene Ontology Term ID||Qualifier||Aspect||Method||Evidence||Source||Assigned On||Annotation Extension||Reference|
|Evidence ID||Analyze ID||Gene||Gene Systematic Name||Phenotype||Experiment Type||Experiment Type Category||Mutant Information||Strain Background||Chemical||Details||Reference|
|Evidence ID||Analyze ID||Regulator||Regulator Systematic Name||Target||Target Systematic Name||Experiment||Assay||Construct||Conditions||Strain Background||Reference|