title: Protein kinase inhibitors can suppress stress-induced dissociation of Hsp27.
abstract: We previously showed that the aggregated form of Hsp27 in cultured cells becomes dissociated as a result of phosphorylation with various types of stress. In order to clarify the signal transduction cascade involved, the effects of various inhibitors of protein kinases and dithiothreitol on the dissociation of Hsp27 were here examined by means of an immunoassay after fractionation of cell extracts by sucrose density gradient centrifugation. The dissociation of Hsp27 induced by exposure of U251 MG human glioma cells to metals (NaAsO2 and CdCl2), hypertonic stress (sorbitol and NaCI), or anisomycin, an activator of p38 mitogen-activated protein (MAP) kinase, was completely suppressed by the presence of SB 203580 or PD 169316, inhibitors of p38 MAP kinase, but not by PD 98059 and Uo 126, inhibitors of MAP kinase kinase (MEK), nor by staurosporine, Go 6983, and bisindolylmaleimide I, inhibitors of protein kinase C. Phorbol ester (PMA)-induced dissociation of Hsp27 was completely suppressed by staurosporine, Go 6983, or bisindolylmaleimide I and partially suppressed by SB 203580, or PD 169316 but not by PD 98059 or Uo 126, indicating mediation by 2 cascades. The presence of 1 mM dithiothreitol in the culture medium during exposure to chemicals suppressed the dissociation of Hsp27 induced by arsenite and CdCl2 but not by other chemicals. These results suggest that the phosphorylation of Hsp27 is catalyzed by 2 protein kinases, p38 MAP kinase-activated protein (MAPKAP) kinase-2/3 and protein kinase C. In addition, metal-induced signals are sensitive to reducing power.
authors.affiliation: Department of Biochemistry, Institute for Developmental Research, Aichi Human Service Center, Kasugai, Japan. kato@inst-hsc.pref.aichi.jp
authors.forename: K
authors.initials: K
authors.lastname: Kato
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: I
authors.initials: I
authors.lastname: Iwamoto
authors.forename: K
authors.initials: K
authors.lastname: Lida
authors.forename: Y
authors.initials: Y
authors.lastname: Inaguma
mesh_terms: D000841:Anisomycin / Q000494:pharmacology; D018053:Arsenites / Q000494:pharmacology; D002070:Butadienes / Q000494:pharmacology; D019256:Cadmium Chloride / Q000494:pharmacology; D002273:Carcinogens / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005419:Flavonoids / Q000494:pharmacology; D005910:Glioma; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007093:Imidazoles / Q000494:pharmacology*; D020930:MAP Kinase Kinase Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology; D009997:Osmotic Pressure / Q000187:drug effects*; D016877:Oxidants / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011725:Pyridines / Q000494:pharmacology*; D017670:Sodium Compounds / Q000494:pharmacology; D013012:Sorbitol / Q000494:pharmacology; D019311:Staurosporine / Q000494:pharmacology; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D048051:p38 Mitogen-Activated Protein Kinases
chemical_list: D018053:Arsenites; D002070:Butadienes; D002273:Carcinogens; D004791:Enzyme Inhibitors; D005419:Flavonoids; D006360:Heat-Shock Proteins; D007093:Imidazoles; D009570:Nitriles; D016877:Oxidants; D011500:Protein Synthesis Inhibitors; D011725:Pyridines; D017670:Sodium Compounds; C113580:U 0126; C017947:sodium arsenite; D013012:Sorbitol; D000841:Anisomycin; D006861:Hydrogen Peroxide; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D020930:MAP Kinase Kinase Kinases; C408604:2-(4-nitrophenyl)-4-(4-fluorophenyl)-5-(4-pyridinyl)-1H-imidazole; D019311:Staurosporine; D019256:Cadmium Chloride; D013755:Tetradecanoylphorbol Acetate; C093642:SB 203580; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D004229:Dithiothreitol