title: 1,4-Naphthoquinone activates the HSP90/HSF1 pathway through the S-arylation of HSP90 in A431 cells: Negative regulation of the redox signal transduction pathway by persulfides/polysulfides.
abstract: The current consensus is that environmental electrophiles activate redox signal transduction pathways through covalent modification of sensor proteins with reactive thiol groups at low concentrations, while they cause cell damage at higher concentrations. We previously exposed human carcinoma A431 cells to the atmospheric electrophile 1,4-naphthoquinone (1,4-NQ) and found that heat shock protein 90 (HSP90), a negative regulator of heat shock factor 1 (HSF1), was a target of 1,4-NQ. In the study presented here, we determined whether 1,4-NQ activates HSF1. We also examined whether such redox signaling could be regulated by nucleophilic sulfur species. Exposure of A431 cells to 1,4-NQ covalently modified cellular HSP90, resulting in repression of the association between HSF1 with HSP90, thereby enhancing HSF1 translocation into the nuclei. Liquid chromatography-tandem mass spectrometry analysis with recombinant HSP90 revealed that the modifications site were Cys412 and Cys564. We found that HSF1 activation mediated by 1,4-NQ upregulated downstream genes, such as HSPA6. HSF1 knockdown accelerated 1,4-NQ-mediated cytotoxicity in the cells. While simultaneous treatment with reactive persulfide and polysulfide, Na2S2and Na2S4, blocked 1,4-NQ-dependent protein modification and HSF1 activation in A431 cells, the knockdown of Cys persulfide producing enzymes cystathionine β-synthase (CBS) and/or cystathionine γ-lyase (CSE) enhanced these phenomena. 1,4-NQ-thiol adduct and 1,4-NQ-S-1,4-NQ adduct were produced during the enzymatic reaction of recombinant CSE in the presence of 1,4-NQ. The results suggest that activation of the HSP90-HSF1 signal transduction pathway mediated by 1,4-NQ protects cells against 1,4-NQ and that per/polysulfides can diminish the reactivity of 1,4-NQ by forming sulfur adducts.
authors.affiliation: Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan; Doctoral Program in Biomedical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Abiko
authors.affiliation: Leading Graduate School Doctoral Program, Ph.D. Program in Human Biology, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Liang
authors.initials: L
authors.lastname: Sha
authors.affiliation: Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan; Doctoral Program in Biomedical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan; Leading Graduate School Doctoral Program, Ph.D. Program in Human Biology, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Shinkai
authors.affiliation: Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Takamitsu
authors.initials: T
authors.lastname: Unoki
authors.affiliation: Doctoral Program in Biomedical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Nho Cong
authors.initials: NC
authors.lastname: Luong
authors.affiliation: Laboratory of Pharmacology, Showa Pharmaceutical University, Tokyo 194-8543, Japan.
authors.forename: Yukihiro
authors.initials: Y
authors.lastname: Tsuchiya
authors.affiliation: Laboratory of Pharmacology, Showa Pharmaceutical University, Tokyo 194-8543, Japan.
authors.forename: Yasuo
authors.initials: Y
authors.lastname: Watanabe
authors.affiliation: Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Hirose
authors.affiliation: Department of Environmental Health Sciences and Molecular Toxicology, Tohoku University Graduate School of Medicine, Sendai, Miyagi 980-8575, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Akaike
authors.affiliation: Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan; Doctoral Program in Biomedical Sciences, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan; Leading Graduate School Doctoral Program, Ph.D. Program in Human Biology, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan. Electronic address: yk-em-tu@md.tsukuba.ac.jp.
authors.forename: Yoshito
authors.initials: Y
authors.lastname: Kumagai
mesh_terms: D045744:Cell Line, Tumor; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D002853:Chromatography, Liquid; D003545:Cysteine / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics*; D018841:HSP90 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D000076249:Heat Shock Transcription Factors / Q000235:genetics*; D006801:Humans; D009285:Naphthoquinones / Q000008:administration & dosage* / Q000378:metabolism; D010084:Oxidation-Reduction; D015398:Signal Transduction / Q000187:drug effects; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism; D053719:Tandem Mass Spectrometry
chemical_list: D004268:DNA-Binding Proteins; C000619706:HSF1 protein, human; D018840:HSP70 Heat-Shock Proteins; D018841:HSP90 Heat-Shock Proteins; C544598:HSPA6 protein, human; D000076249:Heat Shock Transcription Factors; D009285:Naphthoquinones; D013438:Sulfhydryl Compounds; D013455:Sulfur; D003545:Cysteine; C035342:1,4-naphthoquinone
keywords: 1,4-Naphthoquinone; Electrophile; Heat shock factor 1; Heat shock protein 90; Persulfides; Polysulfides