气候变化领域集成服务门户(CCPortal): High-throughput suppressor screen demonstrates that RcsF monitors outer membrane integrity and not Bam complex function

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DOI	10.1073/pnas.2100369118
	High-throughput suppressor screen demonstrates that RcsF monitors outer membrane integrity and not Bam complex function
	Tata M.; Kumar S.; Lach S.R.; Saha S.; Hart E.M.; Konovalova A.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:32
英文摘要	The regulator of capsule synthesis (Rcs) is a complex signaling cascade that monitors gram-negative cell envelope integrity. The outer membrane (OM) lipoprotein RcsF is the sensory component, but how RcsF functions remains elusive. RcsF interacts with the β-barrel assembly machinery (Bam) complex, which assembles RcsF in complex with OM proteins (OMPs), resulting in RcsF's partial cell surface exposure. Elucidating whether RcsF/Bam or RcsF/OMP interactions are important for its sensing function is challenging because the Bam complex is essential, and partial loss-of-function mutations broadly compromise the OM biogenesis. Our recent discovery that, in the absence of nonessential component BamE, RcsF inhibits function of the central component BamA provided a genetic tool to select mutations that specifically prevent RcsF/BamA interactions. We employed a high-throughput suppressor screen to isolate a collection of such rcsF and bamA mutants and characterized their impact on RcsF/OMP assembly and Rcs signaling. Using these mutants and BamA inhibitors MRL-494L and darobactin, we provide multiple lines of evidence against the model in which RcsF senses Bam complex function. We show that Rcs activation in bam mutants results from secondary OM and lipopolysaccharide defects and that RcsF/OMP assembly is required for this activation, supporting an active role of RcsF/OMP complexes in sensing OM stress. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Envelope biogenesis; Envelope stress response; Rcs phosphorelay; Surface-exposed lipoproteins
语种	英语
scopus关键词	BamA protein, E coli; BamB protein, E coli; BamE protein, E coli; darobactin; Escherichia coli protein; lipopolysaccharide; multiprotein complex; outer membrane protein; phenylpropionic acid derivative; rcsF protein, E coli; bacterial capsule; chemistry; gene expression regulation; genetics; high throughput screening; metabolism; mutation; suppressor gene; Bacterial Capsules; Bacterial Outer Membrane Proteins; Escherichia coli Proteins; Gene Expression Regulation, Bacterial; Genes, Suppressor; High-Throughput Screening Assays; Lipopolysaccharides; Multiprotein Complexes; Mutation; Phenylpropionates
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/251080
作者单位	Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, TX 77030, United States; MD Anderson UTHealth Graduate School of Biomedical Sciences, Houston, TX 77030, United States; Department of Molecular Biology, Princeton University, Princeton, NJ 08540, United States; Department of Microbiology, Harvard Medical School, Boston, MA 02115, United States
推荐引用方式 GB/T 7714	Tata M.,Kumar S.,Lach S.R.,et al. High-throughput suppressor screen demonstrates that RcsF monitors outer membrane integrity and not Bam complex function[J],2021,118(32).
APA	Tata M.,Kumar S.,Lach S.R.,Saha S.,Hart E.M.,&Konovalova A..(2021).High-throughput suppressor screen demonstrates that RcsF monitors outer membrane integrity and not Bam complex function.Proceedings of the National Academy of Sciences of the United States of America,118(32).
MLA	Tata M.,et al."High-throughput suppressor screen demonstrates that RcsF monitors outer membrane integrity and not Bam complex function".Proceedings of the National Academy of Sciences of the United States of America 118.32(2021).