<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release

ABSTRACT Enterococcus faecium is an important multidrug-resistant nosocomial pathogen causing biofilm-mediated infections in patients with medical devices. Insight into E. faecium biofilm pathogenesis is pivotal for the development of new strategies to prevent and treat these infections. In several...

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Autores principales:	Fernanda L. Paganelli, Rob J. L. Willems, Pamela Jansen, Antoni Hendrickx, Xinglin Zhang, Marc J. M. Bonten, Helen L. Leavis
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2013
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/7e0816184cec4873905426f3d11ca8a9
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spelling	oai:doaj.org-article:7e0816184cec4873905426f3d11ca8a92021-11-15T15:40:29Z<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release10.1128/mBio.00154-132150-7511https://doaj.org/article/7e0816184cec4873905426f3d11ca8a92013-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00154-13https://doaj.org/toc/2150-7511ABSTRACT Enterococcus faecium is an important multidrug-resistant nosocomial pathogen causing biofilm-mediated infections in patients with medical devices. Insight into E. faecium biofilm pathogenesis is pivotal for the development of new strategies to prevent and treat these infections. In several bacteria, a major autolysin is essential for extracellular DNA (eDNA) release in the biofilm matrix, contributing to biofilm attachment and stability. In this study, we identified and functionally characterized the major autolysin of E. faecium E1162 by a bioinformatic genome screen followed by insertional gene disruption of six putative autolysin genes. Insertional inactivation of locus tag EfmE1162_2692 resulted in resistance to lysis, reduced eDNA release, deficient cell attachment, decreased biofilm, decreased cell wall hydrolysis, and significant chaining compared to that of the wild type. Therefore, locus tag EfmE1162_2692 was considered the major autolysin in E. faecium and renamed atlAEfm. In addition, AtlAEfm was implicated in cell surface exposure of Acm, a virulence factor in E. faecium, and thereby facilitates binding to collagen types I and IV. This is a novel feature of enterococcal autolysins not described previously. Furthermore, we identified (and localized) autolysin-independent DNA release in E. faecium that contributes to cell-cell interactions in the atlAEfm mutant and is important for cell separation. In conclusion, AtlAEfm is the major autolysin in E. faecium and contributes to biofilm stability and Acm localization, making AtlAEfm a promising target for treatment of E. faecium biofilm-mediated infections. IMPORTANCE Nosocomial infections caused by Enterococcus faecium have rapidly increased, and treatment options have become more limited. This is due not only to increasing resistance to antibiotics but also to biofilm-associated infections. DNA is released in biofilm matrix via cell lysis, caused by autolysin, and acts as a matrix stabilizer. In this study, we identified and characterized the major autolysin in E. faecium, which we designated AtlAEfm. atlAEfm disruption resulted in resistance to lysis, reduced extracellular DNA (eDNA), deficient cell attachment, decreased biofilm, decreased cell wall hydrolysis, and chaining. Furthermore, AtlAEfm is associated with Acm cell surface localization, resulting in less binding to collagen types I and IV in the atlAEfm mutant. We also identified AtlAEfm-independent eDNA release that contributes to cell-cell interactions in the atlAEfm mutant. These findings indicate that AtlAEfm is important in biofilm and collagen binding in E. faecium, making AtlAEfm a promising target for treatment of E. faecium infections.Fernanda L. PaganelliRob J. L. WillemsPamela JansenAntoni HendrickxXinglin ZhangMarc J. M. BontenHelen L. LeavisAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 4, Iss 2 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Fernanda L. Paganelli Rob J. L. Willems Pamela Jansen Antoni Hendrickx Xinglin Zhang Marc J. M. Bonten Helen L. Leavis <named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release
description	ABSTRACT Enterococcus faecium is an important multidrug-resistant nosocomial pathogen causing biofilm-mediated infections in patients with medical devices. Insight into E. faecium biofilm pathogenesis is pivotal for the development of new strategies to prevent and treat these infections. In several bacteria, a major autolysin is essential for extracellular DNA (eDNA) release in the biofilm matrix, contributing to biofilm attachment and stability. In this study, we identified and functionally characterized the major autolysin of E. faecium E1162 by a bioinformatic genome screen followed by insertional gene disruption of six putative autolysin genes. Insertional inactivation of locus tag EfmE1162_2692 resulted in resistance to lysis, reduced eDNA release, deficient cell attachment, decreased biofilm, decreased cell wall hydrolysis, and significant chaining compared to that of the wild type. Therefore, locus tag EfmE1162_2692 was considered the major autolysin in E. faecium and renamed atlAEfm. In addition, AtlAEfm was implicated in cell surface exposure of Acm, a virulence factor in E. faecium, and thereby facilitates binding to collagen types I and IV. This is a novel feature of enterococcal autolysins not described previously. Furthermore, we identified (and localized) autolysin-independent DNA release in E. faecium that contributes to cell-cell interactions in the atlAEfm mutant and is important for cell separation. In conclusion, AtlAEfm is the major autolysin in E. faecium and contributes to biofilm stability and Acm localization, making AtlAEfm a promising target for treatment of E. faecium biofilm-mediated infections. IMPORTANCE Nosocomial infections caused by Enterococcus faecium have rapidly increased, and treatment options have become more limited. This is due not only to increasing resistance to antibiotics but also to biofilm-associated infections. DNA is released in biofilm matrix via cell lysis, caused by autolysin, and acts as a matrix stabilizer. In this study, we identified and characterized the major autolysin in E. faecium, which we designated AtlAEfm. atlAEfm disruption resulted in resistance to lysis, reduced extracellular DNA (eDNA), deficient cell attachment, decreased biofilm, decreased cell wall hydrolysis, and chaining. Furthermore, AtlAEfm is associated with Acm cell surface localization, resulting in less binding to collagen types I and IV in the atlAEfm mutant. We also identified AtlAEfm-independent eDNA release that contributes to cell-cell interactions in the atlAEfm mutant. These findings indicate that AtlAEfm is important in biofilm and collagen binding in E. faecium, making AtlAEfm a promising target for treatment of E. faecium infections.
format	article
author	Fernanda L. Paganelli Rob J. L. Willems Pamela Jansen Antoni Hendrickx Xinglin Zhang Marc J. M. Bonten Helen L. Leavis
author_facet	Fernanda L. Paganelli Rob J. L. Willems Pamela Jansen Antoni Hendrickx Xinglin Zhang Marc J. M. Bonten Helen L. Leavis
author_sort	Fernanda L. Paganelli
title	<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release
title_short	<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release
title_full	<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release
title_fullStr	<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release
title_full_unstemmed	<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release
title_sort	<named-content content-type="genus-species">enterococcus faecium</named-content> biofilm formation: identification of major autolysin atla<sub>efm</sub>, associated acm surface localization, and atla<sub>efm</sub>-independent extracellular dna release
publisher	American Society for Microbiology
publishDate	2013
url	https://doaj.org/article/7e0816184cec4873905426f3d11ca8a9
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<named-content content-type="genus-species">Enterococcus faecium</named-content> Biofilm Formation: Identification of Major Autolysin AtlA<sub>Efm</sub>, Associated Acm Surface Localization, and AtlA<sub>Efm</sub>-Independent Extracellular DNA Release

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