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Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile

Cell wall glycopolymers (CWPGs) in Gram-positive bacteria have been reported to be involved in several bacterial processes. These polymers, pillars for proteins and S-layer, are essential for the bacterial surface setup, could be essential for growth, and, in pathogens, participate most often in vir...

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Autores principales: Malet-Villemagne, Jeanne, Yucheng, Liang, Evanno, Laurent, Denis-Quanquin, Sandrine, Hugonnet, Jean-Emmanuel, Arthur, Michel, Janoir, Claire, Candela, Thomas
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Microbiology 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10100865/
https://www.ncbi.nlm.nih.gov/pubmed/36815772
http://dx.doi.org/10.1128/spectrum.04227-22
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author Malet-Villemagne, Jeanne
Yucheng, Liang
Evanno, Laurent
Denis-Quanquin, Sandrine
Hugonnet, Jean-Emmanuel
Arthur, Michel
Janoir, Claire
Candela, Thomas
author_facet Malet-Villemagne, Jeanne
Yucheng, Liang
Evanno, Laurent
Denis-Quanquin, Sandrine
Hugonnet, Jean-Emmanuel
Arthur, Michel
Janoir, Claire
Candela, Thomas
author_sort Malet-Villemagne, Jeanne
collection PubMed
description Cell wall glycopolymers (CWPGs) in Gram-positive bacteria have been reported to be involved in several bacterial processes. These polymers, pillars for proteins and S-layer, are essential for the bacterial surface setup, could be essential for growth, and, in pathogens, participate most often in virulence. CWGPs are covalently anchored to peptidoglycan by proteins that belong to the LytR-CpsA-PSr (LCP) family. This anchoring, important for growth, was reported as essential for some bacteria such as Bacillus subtilis, but the reason why CWGP anchoring is essential remains unknown. We studied LcpA and LcpB of Clostridioides difficile and showed that they have a redundant activity. To delete both lcp genes, we set up the first conditional-lethal mutant method in C. difficile and showed that polysaccharide II (PSII) anchoring at the bacterial surface is essential for C. difficile survival. In the conditional-lethal mutant, C. difficile morphology was impaired, suggesting that peptidoglycan synthesis was affected. Because Lcp proteins are transferring CWPGs from the C(55)-undecaprenyl phosphate (also needed in the peptidoglycan synthesis process), we assumed that there was competition between PSII and peptidoglycan synthesis pathways. We confirmed that UDP-MurNAc-pentapeptide precursor was accumulated, showing that peptidoglycan synthesis was blocked. Our results provide an explanation for the essentiality of PSII anchoring in C. difficile and suggest that the essentiality of the anchoring of CWPGs in other bacteria can also be explained by the blocking of peptidoglycan synthesis. To conclude, our results suggest that Lcps are potential new targets to combat C. difficile infection. IMPORTANCE Cell wall glycopolymers (CWGPs) in Gram-positive bacteria have been reported to be involved in several bacterial processes. CWGP anchoring to peptidoglycan is important for growth and virulence. We set up the first conditional-lethal mutant method in Clostridioides difficile to study LcpA and LcpB involved in the anchoring of CWPGs to peptidoglycan. This study offers new tools to reveal the role of essential genes in C. difficile. LcpA and LcpB activity was shown to be essential, suggesting that they are potential new targets to combat C. difficile infection. In this study, we also showed that there is competition between the polysaccharide II synthesis pathway and peptidoglycan synthesis that probably exists in other Gram-positive bacteria. A better understanding of these mechanisms allows us to define the Lcp proteins as a therapeutic target for potential design of novel antibiotics against pathogenic Gram-positive bacteria.
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spelling pubmed-101008652023-04-14 Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile Malet-Villemagne, Jeanne Yucheng, Liang Evanno, Laurent Denis-Quanquin, Sandrine Hugonnet, Jean-Emmanuel Arthur, Michel Janoir, Claire Candela, Thomas Microbiol Spectr Research Article Cell wall glycopolymers (CWPGs) in Gram-positive bacteria have been reported to be involved in several bacterial processes. These polymers, pillars for proteins and S-layer, are essential for the bacterial surface setup, could be essential for growth, and, in pathogens, participate most often in virulence. CWGPs are covalently anchored to peptidoglycan by proteins that belong to the LytR-CpsA-PSr (LCP) family. This anchoring, important for growth, was reported as essential for some bacteria such as Bacillus subtilis, but the reason why CWGP anchoring is essential remains unknown. We studied LcpA and LcpB of Clostridioides difficile and showed that they have a redundant activity. To delete both lcp genes, we set up the first conditional-lethal mutant method in C. difficile and showed that polysaccharide II (PSII) anchoring at the bacterial surface is essential for C. difficile survival. In the conditional-lethal mutant, C. difficile morphology was impaired, suggesting that peptidoglycan synthesis was affected. Because Lcp proteins are transferring CWPGs from the C(55)-undecaprenyl phosphate (also needed in the peptidoglycan synthesis process), we assumed that there was competition between PSII and peptidoglycan synthesis pathways. We confirmed that UDP-MurNAc-pentapeptide precursor was accumulated, showing that peptidoglycan synthesis was blocked. Our results provide an explanation for the essentiality of PSII anchoring in C. difficile and suggest that the essentiality of the anchoring of CWPGs in other bacteria can also be explained by the blocking of peptidoglycan synthesis. To conclude, our results suggest that Lcps are potential new targets to combat C. difficile infection. IMPORTANCE Cell wall glycopolymers (CWGPs) in Gram-positive bacteria have been reported to be involved in several bacterial processes. CWGP anchoring to peptidoglycan is important for growth and virulence. We set up the first conditional-lethal mutant method in Clostridioides difficile to study LcpA and LcpB involved in the anchoring of CWPGs to peptidoglycan. This study offers new tools to reveal the role of essential genes in C. difficile. LcpA and LcpB activity was shown to be essential, suggesting that they are potential new targets to combat C. difficile infection. In this study, we also showed that there is competition between the polysaccharide II synthesis pathway and peptidoglycan synthesis that probably exists in other Gram-positive bacteria. A better understanding of these mechanisms allows us to define the Lcp proteins as a therapeutic target for potential design of novel antibiotics against pathogenic Gram-positive bacteria. American Society for Microbiology 2023-02-23 /pmc/articles/PMC10100865/ /pubmed/36815772 http://dx.doi.org/10.1128/spectrum.04227-22 Text en Copyright © 2023 Malet-Villemagne et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Research Article
Malet-Villemagne, Jeanne
Yucheng, Liang
Evanno, Laurent
Denis-Quanquin, Sandrine
Hugonnet, Jean-Emmanuel
Arthur, Michel
Janoir, Claire
Candela, Thomas
Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile
title Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile
title_full Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile
title_fullStr Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile
title_full_unstemmed Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile
title_short Polysaccharide II Surface Anchoring, the Achilles’ Heel of Clostridioides difficile
title_sort polysaccharide ii surface anchoring, the achilles’ heel of clostridioides difficile
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10100865/
https://www.ncbi.nlm.nih.gov/pubmed/36815772
http://dx.doi.org/10.1128/spectrum.04227-22
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