Endoribonuclease YbeY Is Essential for RNA Processing and Virulence in Pseudomonas aeruginosa
Endoribonuclease YbeY Is Essential for RNA Processing and Virulence in Pseudomonas aeruginosa
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内切核糖核酸酶 YbeY 对于铜绿假单胞菌的 RNA 加工和毒力至关重要
DOI：
10.1128/mbio.00659-20
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发表时间：
2020-05-01
期刊：
影响因子：
6.4
通讯作者：
Wu, Weihui
中科院分区：
文献类型：
--
作者：
Xia, Yushan;Weng, Yuding;Wu, Weihui
细菌对抗生素的耐药性日益增加，严重威胁着人类的健康。为了开发有效的治疗和预防策略，了解细菌在人体内生长的机制至关重要。转录后调控在细菌适应环境变化中起着重要作用。RNA酶和小RNA是这种调节的关键参与者。在这项研究中，我们证明了RNase YbeY在致病性细菌铜绿假单胞菌的毒力中的关键作用。我们进一步鉴定了小RNA真实的作为YbeY的直接靶点，并阐明了YbeY调控细菌毒力因子表达的途径。我们的研究结果揭示了铜绿假单胞菌复杂的调控网络，并表明YbeY介导的调控途径的推理可能是一种有效的策略，用于开发一种新的治疗策略。摘要转录后调控在病原菌快速适应宿主环境中起着重要作用，而RNA酶通过修饰小分子RNA和mRNA在这一过程中起着关键作用。我们发现，铜绿假单胞菌内切酶YbeY是必需的rRNA加工和细菌的毒力在小鼠急性肺炎模型。转录组学分析表明，敲除ybeY基因导致氧化应激反应基因的下调，包括过氧化氢酶基因katA和katB。一致地，ybeY突变体更容易受到H2O2和嗜中性粒细胞介导的杀伤。katA的过表达恢复了细菌对H2O2和嗜中性粒细胞杀伤的耐受性以及毒力。我们进一步发现，氧化应激反应基因的下调是由于固定相σ因子RpoS的表达缺陷。我们证明了RpoS的自动调节机制，并发现ybeY突变增加了小RNA ReaL的水平，ReaL通过其mRNA的5′ UTR直接抑制rpoS的翻译，随后降低氧化应激反应基因的表达。体外测定证明YbeY直接降解真实的。ybeY突变体中真实的的缺失或rpoS的过表达恢复了细菌对氧化应激的耐受性和毒力。我们还证明了YbeZ结合YbeY，并参与16S rRNA加工和真实的和rpoS的调节以及细菌的毒力。总之，我们的研究结果揭示了YbeY的多效性作用和YbeY介导的调节rpoS通过真实的。重要性细菌抗生素耐药性的增加对人类健康构成严重威胁。为了开发有效的治疗和预防策略，了解细菌在人体内生长的机制至关重要。转录后调控在细菌适应环境变化中起着重要作用。RNA酶和小RNA是这种调节的关键参与者。在这项研究中，我们证明了RNase YbeY在致病性细菌铜绿假单胞菌的毒力中的关键作用。我们进一步鉴定了小RNA真实的作为YbeY的直接靶点，并阐明了YbeY调控细菌毒力因子表达的途径。我们的研究结果揭示了铜绿假单胞菌复杂的调控网络，并表明YbeY介导的调控途径的推理可能是一种有效的策略，用于开发一种新的治疗策略。
The increasing bacterial antibiotic resistance imposes a severe threat to human health. For the development of effective treatment and prevention strategies, it is critical to understand the mechanisms employed by bacteria to grow in the human body. Posttranscriptional regulation plays an important role in bacterial adaptation to environmental changes. RNases and small RNAs are key players in this regulation. In this study, we demonstrate critical roles of the RNase YbeY in the virulence of the pathogenic bacterium Pseudomonas aeruginosa. We further identify the small RNA ReaL as the direct target of YbeY and elucidate the YbeY-regulated pathway on the expression of bacterial virulence factors. Our results shed light on the complex regulatory network of P. aeruginosa and indicate that inference with the YbeY-mediated regulatory pathway might be a valid strategy for the development of a novel treatment strategy. ABSTRACT Posttranscriptional regulation plays an essential role in the quick adaptation of pathogenic bacteria to host environments, and RNases play key roles in this process by modifying small RNAs and mRNAs. We find that the Pseudomonas aeruginosa endonuclease YbeY is required for rRNA processing and the bacterial virulence in a murine acute pneumonia model. Transcriptomic analyses reveal that knocking out the ybeY gene results in downregulation of oxidative stress response genes, including the catalase genes katA and katB. Consistently, the ybeY mutant is more susceptible to H2O2 and neutrophil-mediated killing. Overexpression of katA restores the bacterial tolerance to H2O2 and neutrophil killing as well as virulence. We further find that the downregulation of the oxidative stress response genes is due to defective expression of the stationary-phase sigma factor RpoS. We demonstrate an autoregulatory mechanism of RpoS and find that ybeY mutation increases the level of a small RNA, ReaL, which directly represses the translation of rpoS through the 5′ UTR of its mRNA and subsequently reduces the expression of the oxidative stress response genes. In vitro assays demonstrate direct degradation of ReaL by YbeY. Deletion of reaL or overexpression of rpoS in the ybeY mutant restores the bacterial tolerance to oxidative stress and the virulence. We also demonstrate that YbeZ binds to YbeY and is involved in the 16S rRNA processing and regulation of reaL and rpoS as well as the bacterial virulence. Overall, our results reveal pleiotropic roles of YbeY and the YbeY-mediated regulation of rpoS through ReaL. IMPORTANCE The increasing bacterial antibiotic resistance imposes a severe threat to human health. For the development of effective treatment and prevention strategies, it is critical to understand the mechanisms employed by bacteria to grow in the human body. Posttranscriptional regulation plays an important role in bacterial adaptation to environmental changes. RNases and small RNAs are key players in this regulation. In this study, we demonstrate critical roles of the RNase YbeY in the virulence of the pathogenic bacterium Pseudomonas aeruginosa. We further identify the small RNA ReaL as the direct target of YbeY and elucidate the YbeY-regulated pathway on the expression of bacterial virulence factors. Our results shed light on the complex regulatory network of P. aeruginosa and indicate that inference with the YbeY-mediated regulatory pathway might be a valid strategy for the development of a novel treatment strategy.