Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.

Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging contributor to biofilm-related infections. We recently reported that strains lacking sigma factor B (sigB) in the USA300 lineage of CA-MRSA are unable to develop a biofilm. Interestingly, when spent media from...

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Autores principales:	Megan R Kiedrowski, Jeffrey S Kavanaugh, Cheryl L Malone, Joe M Mootz, Jovanka M Voyich, Mark S Smeltzer, Kenneth W Bayles, Alexander R Horswill
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/8b9996030c7f48c5bee523f30dea8129
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spelling	oai:doaj.org-article:8b9996030c7f48c5bee523f30dea81292021-11-18T07:34:32ZNuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.1932-620310.1371/journal.pone.0026714https://doaj.org/article/8b9996030c7f48c5bee523f30dea81292011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22096493/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging contributor to biofilm-related infections. We recently reported that strains lacking sigma factor B (sigB) in the USA300 lineage of CA-MRSA are unable to develop a biofilm. Interestingly, when spent media from a USA300 sigB mutant was incubated with other S. aureus strains, biofilm formation was inhibited. Following fractionation and mass spectrometry analysis, the major anti-biofilm factor identified in the spent media was secreted thermonuclease (Nuc). Considering reports that extracellular DNA (eDNA) is an important component of the biofilm matrix, we investigated the regulation and role of Nuc in USA300. The expression of the nuc gene was increased in a sigB mutant, repressed by glucose supplementation, and was unaffected by the agr quorum-sensing system. A FRET assay for Nuc activity was developed and confirmed the regulatory results. A USA300 nuc mutant was constructed and displayed an enhanced biofilm-forming capacity, and the nuc mutant also accumulated more high molecular weight eDNA than the WT and regulatory mutant strains. Inactivation of nuc in the USA300 sigB mutant background partially repaired the sigB biofilm-negative phenotype, suggesting that nuc expression contributes to the inability of the mutant to form biofilm. To test the generality of the nuc mutant biofilm phenotypes, the mutation was introduced into other S. aureus genetic backgrounds and similar increases in biofilm formation were observed. Finally, using multiple S. aureus strains and regulatory mutants, an inverse correlation between Nuc activity and biofilm formation was demonstrated. Altogether, our findings confirm the important role for eDNA in the S. aureus biofilm matrix and indicates Nuc is a regulator of biofilm formation.Megan R KiedrowskiJeffrey S KavanaughCheryl L MaloneJoe M MootzJovanka M VoyichMark S SmeltzerKenneth W BaylesAlexander R HorswillPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 11, p e26714 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Megan R Kiedrowski Jeffrey S Kavanaugh Cheryl L Malone Joe M Mootz Jovanka M Voyich Mark S Smeltzer Kenneth W Bayles Alexander R Horswill Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.
description	Community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) is an emerging contributor to biofilm-related infections. We recently reported that strains lacking sigma factor B (sigB) in the USA300 lineage of CA-MRSA are unable to develop a biofilm. Interestingly, when spent media from a USA300 sigB mutant was incubated with other S. aureus strains, biofilm formation was inhibited. Following fractionation and mass spectrometry analysis, the major anti-biofilm factor identified in the spent media was secreted thermonuclease (Nuc). Considering reports that extracellular DNA (eDNA) is an important component of the biofilm matrix, we investigated the regulation and role of Nuc in USA300. The expression of the nuc gene was increased in a sigB mutant, repressed by glucose supplementation, and was unaffected by the agr quorum-sensing system. A FRET assay for Nuc activity was developed and confirmed the regulatory results. A USA300 nuc mutant was constructed and displayed an enhanced biofilm-forming capacity, and the nuc mutant also accumulated more high molecular weight eDNA than the WT and regulatory mutant strains. Inactivation of nuc in the USA300 sigB mutant background partially repaired the sigB biofilm-negative phenotype, suggesting that nuc expression contributes to the inability of the mutant to form biofilm. To test the generality of the nuc mutant biofilm phenotypes, the mutation was introduced into other S. aureus genetic backgrounds and similar increases in biofilm formation were observed. Finally, using multiple S. aureus strains and regulatory mutants, an inverse correlation between Nuc activity and biofilm formation was demonstrated. Altogether, our findings confirm the important role for eDNA in the S. aureus biofilm matrix and indicates Nuc is a regulator of biofilm formation.
format	article
author	Megan R Kiedrowski Jeffrey S Kavanaugh Cheryl L Malone Joe M Mootz Jovanka M Voyich Mark S Smeltzer Kenneth W Bayles Alexander R Horswill
author_facet	Megan R Kiedrowski Jeffrey S Kavanaugh Cheryl L Malone Joe M Mootz Jovanka M Voyich Mark S Smeltzer Kenneth W Bayles Alexander R Horswill
author_sort	Megan R Kiedrowski
title	Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.
title_short	Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.
title_full	Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.
title_fullStr	Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.
title_full_unstemmed	Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.
title_sort	nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/8b9996030c7f48c5bee523f30dea8129
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Nuclease modulates biofilm formation in community-associated methicillin-resistant Staphylococcus aureus.

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