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RNase III CLASH in MRSA uncovers sRNA regulatory networks coupling metabolism to toxin expression
Nature Communications ( IF 16.6 ) Pub Date : 2022-06-22 , DOI: 10.1038/s41467-022-31173-y
Stuart W McKellar 1 , Ivayla Ivanova 1 , Pedro Arede 1 , Rachel L Zapf 2 , Noémie Mercier 3 , Liang-Cui Chu 1 , Daniel G Mediati 4 , Amy C Pickering 5 , Paul Briaud 2 , Robert G Foster 6 , Grzegorz Kudla 6 , J Ross Fitzgerald 5 , Isabelle Caldelari 3 , Ronan K Carroll 2, 7 , Jai J Tree 4 , Sander Granneman 1
Affiliation  

Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterial pathogen responsible for significant human morbidity and mortality. Post-transcriptional regulation by small RNAs (sRNAs) has emerged as an important mechanism for controlling virulence. However, the functionality of the majority of sRNAs during infection is unknown. To address this, we performed UV cross-linking, ligation, and sequencing of hybrids (CLASH) in MRSA to identify sRNA-RNA interactions under conditions that mimic the host environment. Using a double-stranded endoribonuclease III as bait, we uncovered hundreds of novel sRNA-RNA pairs. Strikingly, our results suggest that the production of small membrane-permeabilizing toxins is under extensive sRNA-mediated regulation and that their expression is intimately connected to metabolism. Additionally, we also uncover an sRNA sponging interaction between RsaE and RsaI. Taken together, we present a comprehensive analysis of sRNA-target interactions in MRSA and provide details on how these contribute to the control of virulence in response to changes in metabolism.



中文翻译:

MRSA 中的 RNase III CLASH 揭示了代谢与毒素表达耦合的 sRNA 调控网络

耐甲氧西林金黄色葡萄球菌(MRSA) 是一种导致人类严重发病和死亡的细菌病原体。小RNA (sRNA) 的转录后调控已成为控制毒力的重要机制。然而,大多数 sRNA 在感染过程中的功能尚不清楚。为了解决这个问题,我们对 MRSA 进行了 UV 交联、连接和杂交测序 (CLASH),以在模拟宿主环境的条件下识别 sRNA-RNA 相互作用。使用双链核糖核酸内切酶 III 作为诱饵,我们发现了数百个新的 sRNA-RNA 对。引人注目的是,我们的结果表明,小膜透化毒素的产生受到 sRNA 介导的广泛调节,并且它们的表达与新陈代谢密切相关。此外,我们还发现了 RsaE 和 RsaI 之间的 sRNA 海绵相互作用。综上所述,我们对 MRSA 中的 sRNA-靶标相互作用进行了全面分析,并提供了这些相互作用如何有助于控制毒力以响应代谢变化的详细信息。

更新日期:2022-06-22
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