气候变化领域集成服务门户(CCPortal): A fluorescence-based genetic screen reveals diverse mechanisms silencing small RNA signaling in E. coli

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DOI	10.1073/pnas.2106964118
	A fluorescence-based genetic screen reveals diverse mechanisms silencing small RNA signaling in E. coli
	Chen J.; To L.; de Mets F.; Luo X.; Majdalani N.; Tai C.-H.; Gottesman S.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:27
英文摘要	As key players of gene regulation in many bacteria, small regulatory RNAs (sRNAs) associated with the RNA chaperone Hfq shape numerous phenotypic traits, including metabolism, stress response and adaptation, as well as virulence. sRNAs can alter target messenger RNA (mRNA) translation and stability via base pairing. sRNA synthesis is generally under tight transcriptional regulation, but other levels of regulation of sRNA signaling are less well understood. Here we used a fluorescence-based functional screen to identify regulators that can quench sRNA signaling of the iron-responsive sRNA RyhB in Escherichia coli. The identified regulators fell into two classes, general regulators (affecting signaling by many sRNAs) and RyhB-specific regulators; we focused on the specific ones here. General regulators include three Hfq-interacting sRNAs, CyaR, ChiX, and McaS, previously found to act through Hfq competition, RNase T, a 3′ to 5′ exonuclease not previously implicated in sRNA degradation, and YhbS, a putative GCN5-related N-acetyltransferase (GNAT). Two specific regulators were identified. AspX, a 3′end-derived small RNA, specifically represses RyhB signaling via an RNA sponging mechanism. YicC, a previously uncharacterized but widely conserved protein, triggers rapid RyhB degradation via collaboration with the exoribonuclease PNPase. These findings greatly expand our knowledge of regulation of bacterial sRNA signaling and suggest complex regulatory networks for controlling iron homeostasis in bacteria. The fluorescence-based genetic screen system described here is a powerful tool expected to accelerate the discovery of novel regulators of sRNA signaling in many bacteria. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Exoribonuclease; Hfq competition; RNA chaperone; RNA sponge; RyhB
语种	英语
scopus关键词	acyltransferase; histone acetyltransferase GCN5; transfer RNA; acyltransferase; bacterial RNA; Escherichia coli protein; fusion protein; ribonuclease; Article; bacterial gene; bacterial growth; bacterial strain; bacterium culture; DNA sequence; enzyme activity; Escherichia coli; fluorescence; gene expression; gene library; gene regulatory network; gene silencing; genetic regulation; genetic screening; genetic transcription; heredity; mobile genetic element; molecular cloning; mRNA expression level; nonhuman; Northern blotting; protein degradation; protein expression; real time polymerase chain reaction; reverse transcription polymerase chain reaction; RNA degradation; RNA interference; RNA metabolism; RNA sequence; RNA sequencing; RNA splicing; RNA stability; RNA translation; sequence analysis; signal transduction; Western blotting; bacterial chromosome; bacterial genome; Escherichia coli; fluorescence; gene expression regulation; genetics; metabolism; plasmid; reporter gene; Acetyltransferases; Chromosomes, Bacterial; Escherichia coli; Escherichia coli Proteins; Fluorescence; Gene Expression Regulation, Bacterial; Gene Silencing; Genes, Reporter; Genetic Testing; Genome, Bacterial; Plasmids; Proteolysis; Recombinant Fusion Proteins; Ribonucleases; RNA, Bacterial; Signal Transduction
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/251138
作者单位	Laboratory of Molecular Biology, National Cancer Institute, Bethesda, MD 20892, United States
推荐引用方式 GB/T 7714	Chen J.,To L.,de Mets F.,et al. A fluorescence-based genetic screen reveals diverse mechanisms silencing small RNA signaling in E. coli[J],2021,118(27).
APA	Chen J..,To L..,de Mets F..,Luo X..,Majdalani N..,...&Gottesman S..(2021).A fluorescence-based genetic screen reveals diverse mechanisms silencing small RNA signaling in E. coli.Proceedings of the National Academy of Sciences of the United States of America,118(27).
MLA	Chen J.,et al."A fluorescence-based genetic screen reveals diverse mechanisms silencing small RNA signaling in E. coli".Proceedings of the National Academy of Sciences of the United States of America 118.27(2021).