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(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...

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Autores principales: 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
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
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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.
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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
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