title: Comparison of the effect of heat shock factor inhibitor, KNK437, on heat shock- and chemical stress-induced hsp30 gene expression in Xenopus laevis A6 cells.
abstract: In this study, we compared the effect of KNK437 (N-formyl-3, 4-methylenedioxy-benzylidene-gamma-butyrolactam), a benzylidene lactam compound, on heat shock and chemical stressor-induced hsp30 gene expression in Xenopus laevis A6 kidney epithelial cells. Previously, KNK437 was shown to inhibit HSE-HSF1 binding activity and heat-induced hsp gene expression. In the present study, Northern and Western blot analysis revealed that pretreatment of A6 cells with KNK437 inhibited hsp30 mRNA and HSP30 and HSP70 protein accumulation induced by chemical stressors including sodium arsenite, cadmium chloride and herbimycin A. In A6 cells subjected to sodium arsenite, cadmium chloride, herbimycin A or a 33 degrees C heat shock treatment, immunocytochemistry and confocal microscopy revealed that HSP30 accumulated primarily in the cytoplasm. However, incubation of A6 cells at 35 degrees C resulted in enhanced HSP30 accumulation in the nucleus. Pre-treatment with 100 microM KNK437 completely inhibited HSP30 accumulation in A6 cells heat shocked at 33 or 35 degrees C as well as cells treated with 10 microM sodium arsenite, 100 microM cadmium chloride or 1 microg/mL herbimycin A. These results show that KNK437 is effective at inhibiting both heat shock- and chemical stress-induced hsp gene expression in amphibian cells.
authors.affiliation: Department of Biology, University of Waterloo, Waterloo, ON, N2L 3G1, Canada.
authors.forename: Janine
authors.initials: J
authors.lastname: Voyer
authors.forename: John J
authors.initials: JJ
authors.lastname: Heikkila
mesh_terms: D000818:Animals; D018053:Arsenites / Q000494:pharmacology; D001559:Benzhydryl Compounds / Q000494:pharmacology*; D016227:Benzoquinones / Q000494:pharmacology; D015152:Blotting, Northern; D019256:Cadmium Chloride / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D050890:HSP30 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006360:Heat-Shock Proteins / Q000037:antagonists & inhibitors*; D018869:Heat-Shock Response / Q000187:drug effects* / Q000502:physiology; D015151:Immunoblotting; D007150:Immunohistochemistry; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D011760:Pyrrolidinones / Q000494:pharmacology*; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D017828:Rifabutin / Q000031:analogs & derivatives; D017670:Sodium Compounds / Q000494:pharmacology; D013312:Stress, Physiological; D013696:Temperature; D013997:Time Factors; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics / Q000502:physiology*
chemical_list: D018053:Arsenites; D001559:Benzhydryl Compounds; D016227:Benzoquinones; D050890:HSP30 Heat-Shock Proteins; D006360:Heat-Shock Proteins; C410027:KNK 437; D047029:Lactams, Macrocyclic; D011760:Pyrrolidinones; D017670:Sodium Compounds; D029867:Xenopus Proteins; C494391:hsp30 protein, Xenopus; D017828:Rifabutin; C017947:sodium arsenite; D012313:RNA; C020754:herbimycin; D019256:Cadmium Chloride