Butanol tolerance regulated by a two-component response regulator Slr1037 in photosynthetic Synechocystis sp. PCC 6803.
Butanol tolerance regulated by a two-component response regulator Slr1037 in photosynthetic Synechocystis sp. PCC 6803.
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DOI：
10.1186/1754-6834-7-89
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发表时间：
2014
影响因子：
6.3
通讯作者：
Zhang W
中科院分区：
文献类型：
--
作者：
Chen L;Wu L;Wang J;Zhang W
丁醇对宿主的高毒性限制了光合蓝藻直接利用CO2生产丁醇。在以前的研究中，我们发现一些双组分信号转导系统（TCSTS）的差异调节集胞藻属PCC 6803丁醇处理后。为了探讨丁醇耐受性的调控机制，本研究通过构建丁醇响应TCSTS基因的基因敲除突变体并进行耐受性分析，发现集胞藻中编码一种新型响应调节因子的孤儿基因slr 1037参与了丁醇耐受性的调控。有趣的是，在测试的几种其他胁迫条件下，Eclslr 1037突变体的生长与野生型相似，这表明其对丁醇耐受性的调节是特异性的。使用定量iTRAQ LC-MS/MS蛋白质组学方法结合实时逆转录PCR，我们进一步确定了可能的丁醇耐受性调节子调控的Slr 1037。结果表明，slr 1037基因缺失后，参与光合作用和糖酵解/糖异生等中枢代谢过程的蛋白质，以及具有胁迫响应功能的谷氧还蛋白、肽蛋氨酸亚砜还原酶和葡萄糖基甘油磷酸合成酶表达下调，表明slr 1037基因可能在对抗丁醇胁迫过程中对多种细胞功能发挥调节作用。这项研究提供了一个蛋白质组学描述的假定丁醇耐受调节子调控slr 1037基因。作为第一个发现的与丁醇耐受性直接相关的信号转导蛋白，响应调节因子Slr 1037有望成为集胞藻丁醇耐受性基因工程的天然候选基因。
Butanol production directly from CO2 in photosynthetic cyanobacteria is restricted by the high toxicity of butanol to the hosts. In previous studies, we have found that a few two-component signal transduction systems (TCSTSs) were differentially regulated in Synechocystis sp. PCC 6803 after butanol treatment. To explore regulatory mechanisms of butanol tolerance, in this work, by constructing gene knockout mutants of the butanol-responsive TCSTS genes and conducting tolerance analysis, we uncovered that an orphan slr1037 gene encoding a novel response regulator was involved in butanol tolerance in Synechocystis. Interestingly, the ∆slr1037 mutant grew similarly to the wild type under several other stress conditions tested, which suggests that its regulation on butanol tolerance is specific. Using a quantitative iTRAQ LC-MS/MS proteomics approach coupled with real-time reverse transcription PCR, we further determined the possible butanol-tolerance regulon regulated by Slr1037. The results showed that, after slr1037 deletion, proteins involved in photosynthesis and glycolysis/gluconeogenesis of central metabolic processes, and glutaredoxin, peptide methionine sulfoxide reductase and glucosylglycerol-phosphate synthase with stress-responsive functions were down-regulated, suggesting that Slr1037 may exhibit regulation to a wide range of cellular functions in combating butanol stress. The study provided a proteomic description of the putative butanol-tolerance regulon regulated by the slr1037 gene. As the first signal transduction protein identified directly related to butanol tolerance, response regulator Slr1037 could be a natural candidate for transcriptional engineering to improve butanol tolerance in Synechocystis.
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