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The δ Subunit of RNA Polymerase Guides Promoter Selectivity and Virulence in Staphylococcus aureus

机译：RNA聚合酶的δ亚基指导金黄色葡萄球菌的启动子选择性和致病性

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In Gram-positive bacteria, and particularly the Firmicutes, the DNA-dependent RNA polymerase (RNAP) complex contains an additional subunit, termed the δ factor, or RpoE. This enigmatic protein has been studied for more than 30 years for various organisms, but its function is still not well understood. In this study, we investigated its role in the major human pathogen Staphylococcus aureus. We showed conservation of important structural regions of RpoE in S. aureus and other species and demonstrated binding to core RNAP that is mediated by the β and/or β′ subunits. To identify the impact of the δ subunit on transcription, we performed transcriptome sequencing (RNA-seq) analysis and observed 191 differentially expressed genes in the rpoE mutant. Ontological analysis revealed, quite strikingly, that many of the downregulated genes were known virulence factors, while several mobile genetic elements (SaPI5 and prophage ?SA3usa) were strongly upregulated. Phenotypically, the rpoE mutant had decreased accumulation and/or activity of a number of key virulence factors, including alpha toxin, secreted proteases, and Panton-Valentine leukocidin (PVL). We further observed significantly decreased survival of the mutant in whole human blood, increased phagocytosis by human leukocytes, and impaired virulence in a murine model of infection. Collectively, our results demonstrate that the δ subunit of RNAP is a critical component of the S. aureus transcription machinery and plays an important role during infection.
机译：在革兰氏阳性细菌，特别是Firmicutes中，DNA依赖性RNA聚合酶（RNAP）复合物包含一个额外的亚基，称为δ因子或RpoE。这种神秘的蛋白质已被各种生物研究了30多年，但其功能仍不为人所知。在这项研究中，我们调查了其在主要人类病原体金黄色葡萄球菌中的作用。我们显示了金黄色葡萄球菌和其他物种中RpoE的重要结构区域的保守性，并证明了与由β和/或β'亚基介导的核心RNAP的结合。为了确定δ亚基对转录的影响，我们进行了转录组测序（RNA-seq）分析，并观察到rpoE突变体中的191个差异表达基因。令人吃惊的是，本体分析表明，许多被下调的基因是已知的毒力因子，而一些流动遗传元件（SaPI5和Prohage？SA3usa）则被上调。从表型上看，rpoE突变体的许多关键毒力因子（包括α毒素，分泌的蛋白酶和Panton-Valentine leukocidin（PVL））的积累和/或活性降低。我们进一步观察到突变体在全人类血液中的存活率显着降低，人类白细胞的吞噬作用增加，并且在鼠类感染模型中毒力受损。总的来说，我们的结果表明RNAP的δ亚基是金黄色葡萄球菌转录机制的关键组成部分，并且在感染过程中起着重要的作用。

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