Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus

Megan R. Kiedrowski, Jeffrey S. Kavanaugh, Cheryl L. Malone, Joe M. Mootz, Jovanka M. Voyich, Mark S. Smeltzer, Kenneth W Bayles, Alexander R. Horswill

Research output: Contribution to journalArticle

111 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Article numbere26714
JournalPloS one
Volume6
Issue number11
DOIs
StatePublished - Nov 11 2011

Fingerprint

Methicillin
nucleases
Biofilms
Methicillin-Resistant Staphylococcus aureus
biofilm
sigma factors
Sigma Factor
mutants
Staphylococcus aureus
methicillin-resistant Staphylococcus aureus
DNA
micrococcal nuclease
Micrococcal Nuclease
Phenotype
phenotype
Quorum Sensing
quorum sensing
Fractionation
genetic background
Mass spectrometry

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Kiedrowski, M. R., Kavanaugh, J. S., Malone, C. L., Mootz, J. M., Voyich, J. M., Smeltzer, M. S., ... Horswill, A. R. (2011). Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus. PloS one, 6(11), [e26714]. https://doi.org/10.1371/journal.pone.0026714

Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus. / Kiedrowski, Megan R.; Kavanaugh, Jeffrey S.; Malone, Cheryl L.; Mootz, Joe M.; Voyich, Jovanka M.; Smeltzer, Mark S.; Bayles, Kenneth W; Horswill, Alexander R.

In: PloS one, Vol. 6, No. 11, e26714, 11.11.2011.

Research output: Contribution to journalArticle

Kiedrowski, MR, Kavanaugh, JS, Malone, CL, Mootz, JM, Voyich, JM, Smeltzer, MS, Bayles, KW & Horswill, AR 2011, 'Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus', PloS one, vol. 6, no. 11, e26714. https://doi.org/10.1371/journal.pone.0026714
Kiedrowski MR, Kavanaugh JS, Malone CL, Mootz JM, Voyich JM, Smeltzer MS et al. Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus. PloS one. 2011 Nov 11;6(11). e26714. https://doi.org/10.1371/journal.pone.0026714
Kiedrowski, Megan R. ; Kavanaugh, Jeffrey S. ; Malone, Cheryl L. ; Mootz, Joe M. ; Voyich, Jovanka M. ; Smeltzer, Mark S. ; Bayles, Kenneth W ; Horswill, Alexander R. / Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus. In: PloS one. 2011 ; Vol. 6, No. 11.
@article{2931631e2f034ad3afb3a01a0ae4b1d0,
title = "Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus",
abstract = "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.",
author = "Kiedrowski, {Megan R.} and Kavanaugh, {Jeffrey S.} and Malone, {Cheryl L.} and Mootz, {Joe M.} and Voyich, {Jovanka M.} and Smeltzer, {Mark S.} and Bayles, {Kenneth W} and Horswill, {Alexander R.}",
year = "2011",
month = "11",
day = "11",
doi = "10.1371/journal.pone.0026714",
language = "English (US)",
volume = "6",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "11",

}

TY - JOUR

T1 - Nuclease modulates biofilm formation in community-associated methicillin-resistant staphylococcus aureus

AU - Kiedrowski, Megan R.

AU - Kavanaugh, Jeffrey S.

AU - Malone, Cheryl L.

AU - Mootz, Joe M.

AU - Voyich, Jovanka M.

AU - Smeltzer, Mark S.

AU - Bayles, Kenneth W

AU - Horswill, Alexander R.

PY - 2011/11/11

Y1 - 2011/11/11

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=80855133573&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=80855133573&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0026714

DO - 10.1371/journal.pone.0026714

M3 - Article

C2 - 22096493

AN - SCOPUS:80855133573

VL - 6

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 11

M1 - e26714

ER -