title: Intramolecular repression of mouse heat shock factor 1.
abstract: The pathway leading to transcriptional activation of heat shock genes involves a step of heat shock factor 1 (HSF1) trimerization required for high-affinity binding of this activator protein to heat shock elements (HSEs) in the promoters. Previous studies have shown that in vivo the trimerization is negatively regulated at physiological temperatures by a mechanism that requires multiple hydrophobic heptad repeats (HRs) which may form a coiled coil in the monomer. To investigate the minimal requirements for negative regulation, in this work we have examined mouse HSF1 translated in rabbit reticulocyte lysate or extracted from Escherichia coli after limited expression. We show that under these conditions HSF1 behaves as a monomer which can be induced by increases in temperature to form active HSE-binding trimers and that mutations of either HR region cause activation in both systems. Furthermore, temperature elevations and acidic buffers activate purified HSF1, and mild proteolysis excises fragments which form HSE-binding oligomers. These results suggest that oligomerization can be repressed in the monomer, as previously proposed, and that repression can be relieved in the apparent absence of regulatory proteins. An intramolecular mechanism may be central for the regulation of this transcription factor in mammalian cells, although not necessarily sufficient.
authors.affiliation: Biological and Technological Research Department (DIBIT), San Raffaele Scientific Institute, Milan, Italy.
authors.forename: T
authors.initials: T
authors.lastname: Farkas
authors.forename: Y A
authors.initials: YA
authors.lastname: Kutskova
authors.forename: V
authors.initials: V
authors.lastname: Zimarino
mesh_terms: D000818:Animals; D007611:Aprotinin / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors*; D004330:Drosophila; D004926:Escherichia coli; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000037:antagonists & inhibitors*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051379:Mice; D016296:Mutagenesis; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D014157:Transcription Factors / Q000037:antagonists & inhibitors*; D014981:Xenopus
chemical_list: D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D014157:Transcription Factors; D007611:Aprotinin