Cargando…
(p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli
The outer membrane (OM) protects Gram-negative bacteria from harsh environmental conditions and provides intrinsic resistance to many antimicrobial compounds. The asymmetric OM is characterized by phospholipids in the inner leaflet and lipopolysaccharides (LPS) in the outer leaflet. Previous reports...
Autores principales: | , , , , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10326835/ https://www.ncbi.nlm.nih.gov/pubmed/37426605 http://dx.doi.org/10.1093/femsml/uqad031 |
_version_ | 1785069505646428160 |
---|---|
author | Brückner, Simon Müller, Fabian Schadowski, Laura Kalle, Tyll Weber, Sophia Marino, Emily C Kutscher, Blanka Möller, Anna-Maria Adler, Sabine Begerow, Dominik Steinchen, Wieland Bange, Gert Narberhaus, Franz |
author_facet | Brückner, Simon Müller, Fabian Schadowski, Laura Kalle, Tyll Weber, Sophia Marino, Emily C Kutscher, Blanka Möller, Anna-Maria Adler, Sabine Begerow, Dominik Steinchen, Wieland Bange, Gert Narberhaus, Franz |
author_sort | Brückner, Simon |
collection | PubMed |
description | The outer membrane (OM) protects Gram-negative bacteria from harsh environmental conditions and provides intrinsic resistance to many antimicrobial compounds. The asymmetric OM is characterized by phospholipids in the inner leaflet and lipopolysaccharides (LPS) in the outer leaflet. Previous reports suggested an involvement of the signaling nucleotide ppGpp in cell envelope homeostasis in Escherichia coli. Here, we investigated the effect of ppGpp on OM biosynthesis. We found that ppGpp inhibits the activity of LpxA, the first enzyme of LPS biosynthesis, in a fluorometric in vitro assay. Moreover, overproduction of LpxA resulted in elongated cells and shedding of outer membrane vesicles (OMVs) with altered LPS content. These effects were markedly stronger in a ppGpp-deficient background. We further show that RnhB, an RNase H isoenzyme, binds ppGpp, interacts with LpxA, and modulates its activity. Overall, our study uncovered new regulatory players in the early steps of LPS biosynthesis, an essential process with many implications in the physiology and susceptibility to antibiotics of Gram-negative commensals and pathogens. |
format | Online Article Text |
id | pubmed-10326835 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-103268352023-07-08 (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli Brückner, Simon Müller, Fabian Schadowski, Laura Kalle, Tyll Weber, Sophia Marino, Emily C Kutscher, Blanka Möller, Anna-Maria Adler, Sabine Begerow, Dominik Steinchen, Wieland Bange, Gert Narberhaus, Franz Microlife Research Article The outer membrane (OM) protects Gram-negative bacteria from harsh environmental conditions and provides intrinsic resistance to many antimicrobial compounds. The asymmetric OM is characterized by phospholipids in the inner leaflet and lipopolysaccharides (LPS) in the outer leaflet. Previous reports suggested an involvement of the signaling nucleotide ppGpp in cell envelope homeostasis in Escherichia coli. Here, we investigated the effect of ppGpp on OM biosynthesis. We found that ppGpp inhibits the activity of LpxA, the first enzyme of LPS biosynthesis, in a fluorometric in vitro assay. Moreover, overproduction of LpxA resulted in elongated cells and shedding of outer membrane vesicles (OMVs) with altered LPS content. These effects were markedly stronger in a ppGpp-deficient background. We further show that RnhB, an RNase H isoenzyme, binds ppGpp, interacts with LpxA, and modulates its activity. Overall, our study uncovered new regulatory players in the early steps of LPS biosynthesis, an essential process with many implications in the physiology and susceptibility to antibiotics of Gram-negative commensals and pathogens. Oxford University Press 2023-06-20 /pmc/articles/PMC10326835/ /pubmed/37426605 http://dx.doi.org/10.1093/femsml/uqad031 Text en © The Author(s) 2023. Published by Oxford University Press on behalf of FEMS. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | Research Article Brückner, Simon Müller, Fabian Schadowski, Laura Kalle, Tyll Weber, Sophia Marino, Emily C Kutscher, Blanka Möller, Anna-Maria Adler, Sabine Begerow, Dominik Steinchen, Wieland Bange, Gert Narberhaus, Franz (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli |
title | (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli |
title_full | (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli |
title_fullStr | (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli |
title_full_unstemmed | (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli |
title_short | (p)ppGpp and moonlighting RNases influence the first step of lipopolysaccharide biosynthesis in Escherichia coli |
title_sort | (p)ppgpp and moonlighting rnases influence the first step of lipopolysaccharide biosynthesis in escherichia coli |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10326835/ https://www.ncbi.nlm.nih.gov/pubmed/37426605 http://dx.doi.org/10.1093/femsml/uqad031 |
work_keys_str_mv | AT brucknersimon pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT mullerfabian pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT schadowskilaura pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT kalletyll pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT webersophia pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT marinoemilyc pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT kutscherblanka pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT mollerannamaria pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT adlersabine pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT begerowdominik pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT steinchenwieland pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT bangegert pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli AT narberhausfranz pppgppandmoonlightingrnasesinfluencethefirststepoflipopolysaccharidebiosynthesisinescherichiacoli |