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首页> 外文期刊>BMC Microbiology >Role of sgcR3 in positive regulation of enediyne antibiotic C-1027 production of Streptomyces globisporus C-1027
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Role of sgcR3 in positive regulation of enediyne antibiotic C-1027 production of Streptomyces globisporus C-1027

机译：sgcR3在积极调控烯二炔抗生素C-1027产链霉菌C-1027中的作用

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Background C-1027, produced by Streptomyces globisporus C-1027, is one of the most potent antitumoral agents. The biosynthetic gene cluster of C-1027, previously cloned and sequenced, contains at least three putative regulatory genes, i.e. sgcR1, sgcR2 and sgcR3. The predicted gene products of these genes share sequence similarities to StrR, regulators of AraC/XylS family and TylR. The purpose of this study was to investigate the role of sgcR3 in C-1027 biosynthesis. Results Overexpression of sgcR3 in S. globisporus C-1027 resulted in a 30–40% increase in C-1027 production. Consistent with this, disruption of sgcR3 abolished C-1027 production. Complementation of the sgcR3-disrupted strain R3KO with intact sgcR3 gene could restore C-1027 production. The results from real time RT-PCR analysis in R3KO mutant and wild type strain indicated that not only transcripts of biosynthetic structural genes such as sgcA1 and sgcC4, but also putative regulatory genes, sgcR1 and sgcR2, were significantly decreased in R3KO mutant. The cross-complementation studies showed that sgcR1R2 could functionally complement sgcR3 disruption in trans. Purified N-terminal His10-tagged SgcR3 showed specific DNA-binding activity to the promoter region of sgcR1R2. Conclusion The role of SgcR3 has been proved to be a positive regulator of C-1027 biosynthesis in S. globisporus C-1027. SgcR3 occupies a higher level than SgcR1 and SgcR2 in the regulatory hierarchy that controls C-1027 production and activates the transcription of sgcR1 and sgcR2 by binding directly to the promoter region of sgcR1R2.
机译：背景技术由球形双链霉菌C-1027产生的C-1027是最有效的抗肿瘤剂之一。预先克隆和测序的C-1027的生物合成基因簇含有至少三个推定的调节基因，即sgcR1，sgcR2和sgcR3。这些基因的预测基因产物与StrR，AraC / XylS家族和TylR的调节子具有序列相似性。这项研究的目的是调查sgcR3在C-1027生物合成中的作用。结果globisporus C-1027中sgcR3的过表达导致C-1027产量增加30–40％。与此相一致，对sgcR3的破坏取消了C-1027的生产。完整的sgcR3基因与sgcR3中断的菌株R3KO互补可以恢复C-1027的生产。来自R3KO突变体和野生型菌株的实时RT-PCR分析结果表明，不仅R3KO突变体中的生物合成结构基因如sgcA1和sgcC4的转录本，而且推定的调控基因sgcR1和sgcR2均显着降低。交叉互补研究表明，sgcR1R2可以在功能上互补反式的sgcR3破坏。纯化的带有N末端His 10 标签的SgcR3对sgcR1R2的启动子区域具有特异性的DNA结合活性。结论SgcR3的作用已被证明是球孢链霉菌C-1027中C-1027生物合成的正调节剂。在直接控制sgcR1R2启动子区域的过程中，SgcR3在控制C-1027产生并激活sgcR1和sgcR2转录的调控体系中比SgcR1和SgcR2占据更高的水平。

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