title: Heat Shock Factor 1 Is a Substrate for p38 Mitogen-Activated Protein Kinases.
abstract: Heat shock factor 1 (HSF1) monitors the structural integrity of the proteome. Phosphorylation at S326 is a hallmark for HSF1 activation, but the identity of the kinase(s) phosphorylating this site has remained elusive. We show here that the dietary agent phenethyl isothiocyanate (PEITC) inhibits heat shock protein 90 (Hsp90), the main negative regulator of HSF1; activates p38 mitogen-activated protein kinase (MAPK); and increases S326 phosphorylation, trimerization, and nuclear translocation of HSF1, and the transcription of a luciferase reporter, as well as the endogenous prototypic HSF1 target Hsp70. In vitro, all members of the p38 MAPK family rapidly and stoichiometrically catalyze the S326 phosphorylation. The use of stable knockdown cell lines and inhibitors indicated that among the p38 MAPKs, p38γ is the principal isoform responsible for the phosphorylation of HSF1 at S326 in cells. A protease-mass spectrometry approach confirmed S326 phosphorylation and unexpectedly revealed that p38 MAPK also catalyzes the phosphorylation of HSF1 at S303/307, previously known repressive posttranslational modifications. Thus, we have identified p38 MAPKs as highly efficient catalysts for the phosphorylation of HSF1. Furthermore, our findings suggest that the magnitude and persistence of activation of p38 MAPK are important determinants of the extent and duration of the heat shock response.
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Sharadha
authors.initials: S
authors.lastname: Dayalan Naidu
authors.affiliation: Division of Cardiovascular and Diabetes Medicine, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Calum
authors.initials: C
authors.lastname: Sutherland
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Risco
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Laureano
authors.initials: L
authors.lastname: de la Vega
authors.affiliation: Department of Biology and Biochemistry, University of Bath, Claverton Down, Bath, United Kingdom.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Caunt
authors.affiliation: Division of Signal Transduction Therapy, School of Life Sciences, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: C James
authors.initials: CJ
authors.lastname: Hastie
authors.affiliation: Division of Biological Chemistry and Drug Discovery, School of Life Sciences, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Douglas J
authors.initials: DJ
authors.lastname: Lamont
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Laura
authors.initials: L
authors.lastname: Torrente
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Sudhir
authors.initials: S
authors.lastname: Chowdhry
authors.affiliation: Cardiovascular Center, Medical College of Wisconsin, Milwaukee, Wisconsin, USA.
authors.forename: Ivor J
authors.initials: IJ
authors.lastname: Benjamin
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom.
authors.forename: Stephen M
authors.initials: SM
authors.lastname: Keyse
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología/CSIC, Madrid, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Cuenda
authors.affiliation: Division of Cancer Research, School of Medicine, University of Dundee, Dundee, Scotland, United Kingdom Department of Pharmacology and Molecular Sciences and Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA a.dinkovakostova@dundee.ac.uk.
authors.forename: Albena T
authors.initials: AT
authors.lastname: Dinkova-Kostova
mesh_terms: D000818:Animals; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000235:genetics*; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D018841:HSP90 Heat-Shock Proteins / Q000037:antagonists & inhibitors; D006367:HeLa Cells; D000076249:Heat Shock Transcription Factors; D006801:Humans; D017879:Isothiocyanates / Q000494:pharmacology; D051379:Mice; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D010766:Phosphorylation; D055503:Protein Multimerization; D021381:Protein Transport; D012694:Serine / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; D000076249:Heat Shock Transcription Factors; D017879:Isothiocyanates; D014157:Transcription Factors; D012694:Serine; C058305:phenethyl isothiocyanate; D048051:p38 Mitogen-Activated Protein Kinases