S-Bacillithiolation Protects Against Hypochlorite Stress in Bacillus subtilis as Revealed by Transcriptomics and Redox Proteomics
S-Bacillithiolation Protects Against Hypochlorite Stress in Bacillus subtilis as Revealed by Transcriptomics and Redox Proteomics
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DOI：
10.1074/mcp.m111.009506
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发表时间：
2011-11-01
影响因子：
7
通讯作者：
Antelmann, Haike
中科院分区：
文献类型：
--
作者：
Chi, Bui Khanh;Gronau, Katrin;Antelmann, Haike
蛋白质S-硫醇化是一种翻译后硫醇调节，可控制氧化还原感应转录因子并保护活跃的位点半胱氨酸残基免受不可逆转的氧化。在枯草芽孢杆菌中，通过用氧化还原缓冲杆菌（Cys-Glcn-malate，bSH）形成混合二硫化物，显示出MARR型抑制剂OHRR可感知有机氢过氧化物。在这里，我们研究了由枯草芽孢杆菌中强氧化剂次氯酸引起的转录组和氧化还原蛋白质组的变化。如诱导硫醇和氧化应激特异性SPX，CTSR和PERR调节剂所示，NAOCL应力的表达曲线表示二硫应激。硫代氧化还原蛋白质组学仅鉴定出少数细胞质蛋白，其响应于包括GAPA和Mete在内的NAOCL应激，其硫醇氧化作用。 Shotgun-liquid色谱串联MS分析表明，GAPA，SPX和PERR被NAOCL应力氧化为分子内二硫化物。此外，我们在NaOCL处理的细胞中鉴定了六种S-纤维硫硫化蛋白，包括OHRR抑制剂，两个蛋氨酸合酶Mete和YXJG，无机焦磷酸酶PPAC，3-D-D- D-磷酸甘油酸酯脱氢酶Sera和Putative bacillired yphp。 OHRR抑制剂的S-核硫代化导致OHRA过氧蛋白的上调，该氧化毒素与针对NAOCL的BSH特异性保护在一起。 Mete，YXJG，PPAC和血清的S-纤维硫化会导致次氯酸盐诱导的蛋氨酸饥饿，这是S-Box Regulon的诱导支持的。在大肠杆菌中已经描述了Mete的S-谷胱甘肽化的机理，还导致酶失活和蛋氨酸一个良性营养丰富。总而言之，我们的研究发现，杆菌氧化还原缓冲液在防御次氯酸酸中的重要作用，通过对氧化还原传感调节剂OHRR和蛋氨酸生物合成途径的四个酶的S-核硫代化作用。分子和细胞蛋白质组学10：10.1074/MCP.M111.009506，1-21，2011。
Protein S-thiolation is a post-translational thiol-modification that controls redox-sensing transcription factors and protects active site cysteine residues against irreversible oxidation. In Bacillus subtilis the MarR-type repressor OhrR was shown to sense organic hydroperoxides via formation of mixed disulfides with the redox buffer bacillithiol (Cys-GlcN-Malate, BSH), termed as S-bacillithiolation. Here we have studied changes in the transcriptome and redox proteome caused by the strong oxidant hypochloric acid in B. subtilis. The expression profile of NaOCl stress is indicative of disulfide stress as shown by the induction of the thiol-and oxidative stress-specific Spx, CtsR, and PerR regulons. Thiol redox proteomics identified only few cytoplasmic proteins with reversible thiol-oxidations in response to NaOCl stress that include GapA and MetE. Shotgun-liquid chromatography-tandem MS analyses revealed that GapA, Spx, and PerR are oxidized to intramolecular disulfides by NaOCl stress. Furthermore, we identified six S-bacillithiolated proteins in NaOCl-treated cells, including the OhrR repressor, two methionine synthases MetE and YxjG, the inorganic pyrophosphatase PpaC, the 3-D-phosphoglycerate dehydrogenase SerA, and the putative bacilliredoxin YphP. S-bacillithiolation of the OhrR repressor leads to up-regulation of the OhrA peroxiredoxin that confers together with BSH specific protection against NaOCl. S-bacillithiolation of MetE, YxjG, PpaC and SerA causes hypochlorite-induced methionine starvation as supported by the induction of the S-box regulon. The mechanism of S-glutathionylation of MetE has been described in Escherichia coli also leading to enzyme inactivation and methionine auxotrophy. In summary, our studies discover an important role of the bacillithiol redox buffer in protection against hypochloric acid by S-bacillithiolation of the redox-sensing regulator OhrR and of four enzymes of the methionine biosynthesis pathway. Molecular & Cellular Proteomics 10: 10.1074/mcp.M111.009506, 1-21, 2011.