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全基因组分析 Hfq 结合 RNA 揭示了. 中的铁应答小 RNA RusT。

Genome-wide profiling of Hfq-bound RNAs reveals the iron-responsive small RNA RusT in .

机构信息

Institute of Microbiology, Faculty of Biological Sciences, Friedrich Schiller University, Jena, Germany.

Department of Biology I, Microbiology, Ludwig-Maximilians-University Munich, Munich, Germany.

出版信息

mBio. 2024 Apr 10;15(4):e0315323. doi: 10.1128/mbio.03153-23. Epub 2024 Mar 21.

Abstract

The alphaproteobacterium thrives in oligotrophic environments and is able to optimally exploit minimal resources by entertaining an intricate network of gene expression control mechanisms. Numerous transcriptional activators and repressors have been reported to contribute to these processes, but only few studies have focused on regulation at the post-transcriptional level in . Small RNAs (sRNAs) are a prominent class of regulators of bacterial gene expression, and most sRNAs characterized today engage in direct base-pairing interactions to modulate the translation and/or stability of target mRNAs. In many cases, the ubiquitous RNA chaperone, Hfq, contributes to the establishment of RNA-RNA interactions. Although the deletion of the gene is associated with a severe loss of fitness in , the RNA ligands of the chaperone have remained largely unexplored. Here we report on the identification of coding and non-coding transcripts associated with Hfq in and demonstrate Hfq-dependent post-transcriptional regulation in this organism. We show that the Hfq-bound sRNA RusT is transcriptionally controlled by the NtrYX two-component system and induced in response to iron starvation. By combining RusT pulse expression with whole-genome transcriptome analysis, we determine 16 candidate target transcripts that are deregulated, many of which encode outer membrane transporters. We hence suggest RusT to support remodeling of the cell surface when iron supplies are limited.IMPORTANCEThe conserved RNA-binding protein Hfq contributes significantly to the adaptation of bacteria to different environmental conditions. Hfq not only stabilizes associated sRNAs but also promotes inter-molecular base-pairing interactions with target transcripts. Hfq plays a pivotal role for growth and survival, controlling central metabolism and cell wall synthesis in the oligotroph . However, direct evidence for Hfq-dependent post-transcriptional regulation and potential oligotrophy in has been lacking. Here, we identified sRNAs and mRNAs associated with Hfq , and demonstrated the requirement of Hfq for sRNA-mediated regulation, particularly of outer membrane transporters in .

摘要

α-变形菌在贫营养环境中茁壮成长,能够通过复杂的基因表达控制机制来最优地利用最小的资源。已经报道了许多转录激活因子和阻遏物有助于这些过程,但只有少数研究集中在 中的转录后水平调节。小 RNA (sRNA) 是细菌基因表达调控的主要调控因子之一,目前大多数特征明确的 sRNA 通过直接碱基配对相互作用来调节靶 mRNA 的翻译和/或稳定性。在许多情况下,普遍存在的 RNA 伴侣 Hfq 有助于 RNA-RNA 相互作用的建立。尽管 基因的缺失与 在 中的适应性严重丧失有关,但该伴侣的 RNA 配体在很大程度上仍未得到探索。在这里,我们报告了与 中 Hfq 相关的编码和非编码转录本的鉴定,并证明了该生物体中 Hfq 依赖的转录后调节。我们表明,Hfq 结合的 sRNA RusT 受 NtrYX 双组分系统的转录控制,并在缺铁胁迫下诱导。通过将 RusT 脉冲表达与全基因组转录组分析相结合,我们确定了 16 个候选靶转录本发生了调节,其中许多编码外膜转运蛋白。因此,我们建议 RusT 在铁供应有限时支持 细胞表面的重塑。

重要性

保守的 RNA 结合蛋白 Hfq 对细菌适应不同环境条件有重要贡献。Hfq 不仅稳定相关的 sRNA,还促进与靶转录物的分子间碱基配对相互作用。Hfq 在寡营养 中对生长和存活起着关键作用,控制着中央代谢和细胞壁合成。然而,直接证明 Hfq 依赖的转录后调节和 中的潜在寡营养性的证据一直缺乏。在这里,我们鉴定了与 Hfq 相关的 sRNA 和 mRNA,并证明了 Hfq 对 sRNA 介导的调节,特别是对 的外膜转运蛋白的调节是必需的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2874/11005374/0e58974fc34d/mbio.03153-23.f001.jpg

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