Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase

ABSTRACT Peptidoglycan recycling is a metabolic process by which Gram-negative bacteria reutilize up to half of their cell wall within one generation during vegetative growth. Whether peptidoglycan recycling also occurs in Gram-positive bacteria has so far remained unclear. We show here that three G...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Marina Borisova, Rosmarie Gaupp, Amanda Duckworth, Alexander Schneider, Désirée Dalügge, Maraike Mühleck, Denise Deubel, Sandra Unsleber, Wenqi Yu, Günther Muth, Markus Bischoff, Friedrich Götz, Christoph Mayer
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c65f4220c7e34568b1e914208f42b1e4
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:c65f4220c7e34568b1e914208f42b1e4
record_format	dspace
spelling	oai:doaj.org-article:c65f4220c7e34568b1e914208f42b1e42021-11-15T15:50:15ZPeptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase10.1128/mBio.00923-162150-7511https://doaj.org/article/c65f4220c7e34568b1e914208f42b1e42016-11-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00923-16https://doaj.org/toc/2150-7511ABSTRACT Peptidoglycan recycling is a metabolic process by which Gram-negative bacteria reutilize up to half of their cell wall within one generation during vegetative growth. Whether peptidoglycan recycling also occurs in Gram-positive bacteria has so far remained unclear. We show here that three Gram-positive model organisms, Staphylococcus aureus, Bacillus subtilis, and Streptomyces coelicolor, all recycle the sugar N-acetylmuramic acid (MurNAc) of their peptidoglycan during growth in rich medium. They possess MurNAc-6-phosphate (MurNAc-6P) etherase (MurQ in E. coli) enzymes, which are responsible for the intracellular conversion of MurNAc-6P to N-acetylglucosamine-6-phosphate and d-lactate. By applying mass spectrometry, we observed accumulation of MurNAc-6P in MurNAc-6P etherase deletion mutants but not in either the isogenic parental strains or complemented strains, suggesting that MurQ orthologs are required for the recycling of cell wall-derived MurNAc in these bacteria. Quantification of MurNAc-6P in ΔmurQ cells of S. aureus and B. subtilis revealed small amounts during exponential growth phase (0.19 nmol and 0.03 nmol, respectively, per ml of cells at an optical density at 600 nm [OD600] of 1) but large amounts during transition (0.56 nmol and 0.52 nmol) and stationary (0.53 nmol and 1.36 nmol) phases. The addition of MurNAc to ΔmurQ cultures greatly increased the levels of intracellular MurNAc-6P in all growth phases. The ΔmurQ mutants of S. aureus and B. subtilis showed no growth deficiency in rich medium compared to the growth of the respective parental strains, but intriguingly, they had a severe survival disadvantage in late stationary phase. Thus, although peptidoglycan recycling is apparently not essential for the growth of Gram-positive bacteria, it provides a benefit for long-term survival. IMPORTANCE The peptidoglycan of the bacterial cell wall is turned over steadily during growth. As peptidoglycan fragments were found in large amounts in spent medium of exponentially growing Gram-positive bacteria, their ability to recycle these fragments has been questioned. We conclusively showed recycling of the peptidoglycan component MurNAc in different Gram-positive model organisms and revealed that a MurNAc-6P etherase (MurQ or MurQ ortholog) enzyme is required in this process. We further demonstrated that recycling occurs predominantly during the transition to stationary phase in S. aureus and B. subtilis, explaining why peptidoglycan fragments are found in the medium during exponential growth. We quantified the intracellular accumulation of recycling products in MurNAc-6P etherase gene mutants, revealing that about 5% and 10% of the MurNAc of the cell wall per generation is recycled in S. aureus and B. subtilis, respectively. Importantly, we showed that MurNAc recycling and salvaging does not sustain growth in these bacteria but is used to enhance survival during late stationary phase.Marina BorisovaRosmarie GauppAmanda DuckworthAlexander SchneiderDésirée DalüggeMaraike MühleckDenise DeubelSandra UnsleberWenqi YuGünther MuthMarkus BischoffFriedrich GötzChristoph MayerAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 5 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Marina Borisova Rosmarie Gaupp Amanda Duckworth Alexander Schneider Désirée Dalügge Maraike Mühleck Denise Deubel Sandra Unsleber Wenqi Yu Günther Muth Markus Bischoff Friedrich Götz Christoph Mayer Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase
description	ABSTRACT Peptidoglycan recycling is a metabolic process by which Gram-negative bacteria reutilize up to half of their cell wall within one generation during vegetative growth. Whether peptidoglycan recycling also occurs in Gram-positive bacteria has so far remained unclear. We show here that three Gram-positive model organisms, Staphylococcus aureus, Bacillus subtilis, and Streptomyces coelicolor, all recycle the sugar N-acetylmuramic acid (MurNAc) of their peptidoglycan during growth in rich medium. They possess MurNAc-6-phosphate (MurNAc-6P) etherase (MurQ in E. coli) enzymes, which are responsible for the intracellular conversion of MurNAc-6P to N-acetylglucosamine-6-phosphate and d-lactate. By applying mass spectrometry, we observed accumulation of MurNAc-6P in MurNAc-6P etherase deletion mutants but not in either the isogenic parental strains or complemented strains, suggesting that MurQ orthologs are required for the recycling of cell wall-derived MurNAc in these bacteria. Quantification of MurNAc-6P in ΔmurQ cells of S. aureus and B. subtilis revealed small amounts during exponential growth phase (0.19 nmol and 0.03 nmol, respectively, per ml of cells at an optical density at 600 nm [OD600] of 1) but large amounts during transition (0.56 nmol and 0.52 nmol) and stationary (0.53 nmol and 1.36 nmol) phases. The addition of MurNAc to ΔmurQ cultures greatly increased the levels of intracellular MurNAc-6P in all growth phases. The ΔmurQ mutants of S. aureus and B. subtilis showed no growth deficiency in rich medium compared to the growth of the respective parental strains, but intriguingly, they had a severe survival disadvantage in late stationary phase. Thus, although peptidoglycan recycling is apparently not essential for the growth of Gram-positive bacteria, it provides a benefit for long-term survival. IMPORTANCE The peptidoglycan of the bacterial cell wall is turned over steadily during growth. As peptidoglycan fragments were found in large amounts in spent medium of exponentially growing Gram-positive bacteria, their ability to recycle these fragments has been questioned. We conclusively showed recycling of the peptidoglycan component MurNAc in different Gram-positive model organisms and revealed that a MurNAc-6P etherase (MurQ or MurQ ortholog) enzyme is required in this process. We further demonstrated that recycling occurs predominantly during the transition to stationary phase in S. aureus and B. subtilis, explaining why peptidoglycan fragments are found in the medium during exponential growth. We quantified the intracellular accumulation of recycling products in MurNAc-6P etherase gene mutants, revealing that about 5% and 10% of the MurNAc of the cell wall per generation is recycled in S. aureus and B. subtilis, respectively. Importantly, we showed that MurNAc recycling and salvaging does not sustain growth in these bacteria but is used to enhance survival during late stationary phase.
format	article
author	Marina Borisova Rosmarie Gaupp Amanda Duckworth Alexander Schneider Désirée Dalügge Maraike Mühleck Denise Deubel Sandra Unsleber Wenqi Yu Günther Muth Markus Bischoff Friedrich Götz Christoph Mayer
author_facet	Marina Borisova Rosmarie Gaupp Amanda Duckworth Alexander Schneider Désirée Dalügge Maraike Mühleck Denise Deubel Sandra Unsleber Wenqi Yu Günther Muth Markus Bischoff Friedrich Götz Christoph Mayer
author_sort	Marina Borisova
title	Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase
title_short	Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase
title_full	Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase
title_fullStr	Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase
title_full_unstemmed	Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase
title_sort	peptidoglycan recycling in gram-positive bacteria is crucial for survival in stationary phase
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/c65f4220c7e34568b1e914208f42b1e4
work_keys_str_mv	AT marinaborisova peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT rosmariegaupp peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT amandaduckworth peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT alexanderschneider peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT desireedalugge peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT maraikemuhleck peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT denisedeubel peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT sandraunsleber peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT wenqiyu peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT gunthermuth peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT markusbischoff peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT friedrichgotz peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase AT christophmayer peptidoglycanrecyclingingrampositivebacteriaiscrucialforsurvivalinstationaryphase
_version_	1718427485046046720

Peptidoglycan Recycling in Gram-Positive Bacteria Is Crucial for Survival in Stationary Phase

Ejemplares similares