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The Legionella pneumophila genome evolved to accommodate multiple regulatory mechanisms controlled by the CsrA-system

机译:嗜肺军团菌基因组进化以适应由CsrA系统控制的多种调控机制

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摘要

The carbon storage regulator protein CsrA regulates cellular processes post-transcriptionally by binding to target-RNAs altering translation efficiency and/or their stability. Here we identified and analyzed the direct targets of CsrA in the human pathogen Legionella pneumophila. Genome wide transcriptome, proteome and RNA co-immunoprecipitation followed by deep sequencing of a wild type and a csrA mutant strain identified 479 RNAs with potential CsrA interaction sites located in the untranslated and/or coding regions of mRNAs or of known non-coding sRNAs. Further analyses revealed that CsrA exhibits a dual regulatory role in virulence as it affects the expression of the regulators FleQ, LqsR, LetE and RpoS but it also directly regulates the timely expression of over 40 Dot/Icm substrates. CsrA controls its own expression and the stringent response through a regulatory feedback loop as evidenced by its binding to RelA-mRNA and links it to quorum sensing and motility. CsrA is a central player in the carbon, amino acid, fatty acid metabolism and energy transfer and directly affects the biosynthesis of cofactors, vitamins and secondary metabolites. We describe the first L. pneumophila riboswitch, a thiamine pyrophosphate riboswitch whose regulatory impact is fine-tuned by CsrA, and identified a unique regulatory mode of CsrA, the active stabilization of RNA anti-terminator conformations inside a coding sequence preventing Rho-dependent termination of the gap operon through transcriptional polarity effects. This allows L. pneumophila to regulate the pentose phosphate pathway and the glycolysis combined or individually although they share genes in a single operon. Thus the L. pneumophila genome has evolved to acclimate at least five different modes of regulation by CsrA giving it a truly unique position in its life cycle.
机译:碳储存调节蛋白CsrA通过与靶RNA结合而改变翻译效率和/或稳定性,从而在转录后调节细胞过程。在这里,我们确定并分析了人类病原体肺炎军团菌中CsrA的直接目标。全基因组转录组,蛋白质组和RNA共同免疫沉淀,然后对野生型和csrA突变菌株进行深度测序,鉴定出479个RNA,这些RNA具有潜在的CsrA相互作用位点,位于mRNA或已知非编码sRNA的非翻译和/或编码区域。进一步的分析表明,CsrA在毒力中具有双重调节作用,因为它影响调节剂FleQ,LqsR,LetE和RpoS的表达,但它也直接调节40多种Dot / Icm底物的及时表达。 CsrA通过调节性反馈回路控制其自身的表达和严格的反应,正如其与RelA-mRNA的结合所证明的那样,并将其与群体感应和运动性联系起来。 CsrA是碳,氨基酸,脂肪酸代谢和能量转移的主要参与者,直接影响辅因子,维生素和次生代谢产物的生物合成。我们描述了第一个肺炎链球菌核糖开关,硫胺素焦磷酸核糖开关,其调节作用由CsrA进行了微调,并确定了CsrA的独特调节模式,即在编码序列内部防止Rho依赖性终止的RNA抗终止子构象的主动稳定作用间隙操纵子通过转录极性效应的作用。尽管它们在单个操纵子中共享基因,但是这使肺炎衣原体能够调节戊糖磷酸途径和糖酵解结合起来或单独进行。因此,嗜肺乳杆菌基因组已经进化为适应CsrA的至少五种不同调节模式,使其在其生命周期中处于真正独特的位置。

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