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
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
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DOI：
10.1016/j.freeradbiomed.2016.12.047
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发表时间：
2017-03-01
影响因子：
7.4
通讯作者：
Kumagai, Yoshito
中科院分区：
文献类型：
--
作者：
Abiko, Yumi;Sha, Liang;Kumagai, Yoshito
目前的共识是，环境亲电体激活氧化还原信号转导途径，通过共价修饰的传感器蛋白与反应性硫醇基团在低浓度下，而他们在较高的浓度引起细胞损伤。我们以前将人癌A431细胞暴露于大气亲电试剂1，4-萘醌（1，4-NQ），发现热休克蛋白90（HSP 90），热休克因子1（HSF 1）的负调节剂，是1，4-NQ的靶点。在本研究中，我们确定了1，4-NQ是否激活HSF 1。我们还研究了这种氧化还原信号是否可以调节亲核硫物种。将A431细胞暴露于1，4-NQ共价修饰的细胞HSP 90，导致HSF 1与HSP 90之间的缔合被抑制，从而增强HSF 1向细胞核的易位。液相色谱-串联质谱分析表明，HSP 90的修饰位点为Cys 412和Cys 564。我们发现，由1，4-NQ介导的HSF 1激活上调下游基因，如HSPA 6。HSF 1敲减加速了细胞中1，4-NQ介导的细胞毒性。虽然同时用反应性过硫化物和多硫化物，Na 2S 2和Na 2S 4处理，阻断了A431细胞中1，4-NQ依赖性蛋白质修饰和HSF 1活化，但敲低Cys过硫化物产生酶胱硫醚β-合酶（CBS）和/或胱硫醚γ-裂解酶（CSE）增强了这些现象。在1，4-NQ存在下，在重组CSE的酶促反应期间产生1，4-NQ-硫醇加合物和1，4-NQ-S-1，4-NQ加合物。结果表明，由1，4-NQ介导的HSP 90-HSF 1信号转导通路的激活保护细胞免受1，4-NQ的侵害，并且过/多硫化物可以通过形成硫加合物来降低1，4-NQ的反应性。
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, Na2S2 and Na2S4, blocked 1,4-NQ-dependent protein modification and HSF1 activation in A431 cells, the knockdown of Cys persulfide producing enzymes cystathionine beta-synthase (CBS) and/or cystathionine gamma-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.