Bacillus subtilis as a Platform for Molecular Characterisation of Regulatory Mechanisms of Enterococcus faecalis Resistance against Cell Wall Antibiotics

Publikationstyp:
Zeitschriftenaufsatz
Metadaten:
Autoren
  • Chong Fang
  • Emanuel Stiegeler
  • Gregory M Cook
  • Thorsten Mascher
  • Susanne Gebhard
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000333677500085&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1371/journal.pone.0093169
Externe Identifier
  • Clarivate Analytics Document Solution ID: AE0SW
  • PubMed Identifier: 24676422
ISSN
1932-6203
Ausgabe der Veröffentlichung
3
Zeitschrift
PLOS ONE
Artikelnummer
ARTN e93169
Datum der Veröffentlichung
2014
Status
Published
Titel
<i>Bacillus subtilis</i> as a Platform for Molecular Characterisation of Regulatory Mechanisms of <i>Enterococcus</i> faecalis Resistance against Cell Wall Antibiotics
Sub types
  • Article
Ausgabe der Zeitschrift
9

Datenquelle: Web of Science (Lite)

Andere Metadatenquellen:
Autoren
  • Chong Fang
  • Emanuel Stiegeler
  • Gregory M Cook
  • Thorsten Mascher
  • Susanne Gebhard
DOI
10.1371/journal.pone.0093169
Editoren
  • Willem van Schaik
eISSN
1932-6203
Ausgabe der Veröffentlichung
3
Zeitschrift
PLoS ONE
Sprache
en
Online publication date
2014
Paginierung
e93169 - e93169
Status
Published online
Herausgeber
Public Library of Science (PLoS)
Herausgeber URL
http://dx.doi.org/10.1371/journal.pone.0093169
Datum der Datenerfassung
2022
Titel
Bacillus subtilis as a Platform for Molecular Characterisation of Regulatory Mechanisms of Enterococcus faecalis Resistance against Cell Wall Antibiotics
Ausgabe der Zeitschrift
9

Datenquelle: Crossref

Abstract
To combat antibiotic resistance of Enterococcus faecalis, a better understanding of the molecular mechanisms, particularly of antibiotic detection, signal transduction and gene regulation is needed. Because molecular studies in this bacterium can be challenging, we aimed at exploiting the genetically highly tractable Gram-positive model organism Bacillus subtilis as a heterologous host. Two fundamentally different regulators of E. faecalis resistance against cell wall antibiotics, the bacitracin sensor BcrR and the vancomycin-sensing two-component system VanSB-VanRB, were produced in B. subtilis and their functions were monitored using target promoters fused to reporter genes (lacZ and luxABCDE). The bacitracin resistance system BcrR-BcrAB of E. faecalis was fully functional in B. subtilis, both regarding regulation of bcrAB expression and resistance mediated by the transporter BcrAB. Removal of intrinsic bacitracin resistance of B. subtilis increased the sensitivity of the system. The lacZ and luxABCDE reporters were found to both offer sensitive detection of promoter induction on solid media, which is useful for screening of large mutant libraries. The VanSB-VanRB system displayed a gradual dose-response behaviour to vancomycin, but only when produced at low levels in the cell. Taken together, our data show that B. subtilis is a well-suited host for the molecular characterization of regulatory systems controlling resistance against cell wall active compounds in E. faecalis. Importantly, B. subtilis facilitates the careful adjustment of expression levels and genetic background required for full functionality of the introduced regulators.
Addresses
  • Department Biology I, Microbiology, Ludwig-Maximilians-Universität München, Martinsried, Germany.
Autoren
  • Chong Fang
  • Emanuel Stiegeler
  • Gregory M Cook
  • Thorsten Mascher
  • Susanne Gebhard
DOI
10.1371/journal.pone.0093169
eISSN
1932-6203
Externe Identifier
  • PubMed Identifier: 24676422
  • PubMed Central ID: PMC3968067
Open access
true
ISSN
1932-6203
Ausgabe der Veröffentlichung
3
Zeitschrift
PloS one
Schlüsselwörter
  • Cell Wall
  • Bacillus subtilis
  • Enterococcus faecalis
  • Anti-Bacterial Agents
  • Microbial Sensitivity Tests
  • Transduction, Genetic
  • Drug Resistance, Bacterial
  • Gene Expression
  • Genes, Bacterial
  • Genes, Reporter
  • Promoter Regions, Genetic
Sprache
eng
Medium
Electronic-eCollection
Online publication date
2014
Open access status
Open Access
Paginierung
e93169
Datum der Veröffentlichung
2014
Status
Published
Publisher licence
CC BY
Datum der Datenerfassung
2014
Titel
Bacillus subtilis as a platform for molecular characterisation of regulatory mechanisms of Enterococcus faecalis resistance against cell wall antibiotics.
Sub types
  • Research Support, Non-U.S. Gov't
  • research-article
  • Journal Article
Ausgabe der Zeitschrift
9

Files

https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0093169&type=printable https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24676422/pdf/?tool=EBI https://europepmc.org/articles/PMC3968067?pdf=render

Datenquelle: Europe PubMed Central

Abstract
To combat antibiotic resistance of Enterococcus faecalis, a better understanding of the molecular mechanisms, particularly of antibiotic detection, signal transduction and gene regulation is needed. Because molecular studies in this bacterium can be challenging, we aimed at exploiting the genetically highly tractable Gram-positive model organism Bacillus subtilis as a heterologous host. Two fundamentally different regulators of E. faecalis resistance against cell wall antibiotics, the bacitracin sensor BcrR and the vancomycin-sensing two-component system VanSB-VanRB, were produced in B. subtilis and their functions were monitored using target promoters fused to reporter genes (lacZ and luxABCDE). The bacitracin resistance system BcrR-BcrAB of E. faecalis was fully functional in B. subtilis, both regarding regulation of bcrAB expression and resistance mediated by the transporter BcrAB. Removal of intrinsic bacitracin resistance of B. subtilis increased the sensitivity of the system. The lacZ and luxABCDE reporters were found to both offer sensitive detection of promoter induction on solid media, which is useful for screening of large mutant libraries. The VanSB-VanRB system displayed a gradual dose-response behaviour to vancomycin, but only when produced at low levels in the cell. Taken together, our data show that B. subtilis is a well-suited host for the molecular characterization of regulatory systems controlling resistance against cell wall active compounds in E. faecalis. Importantly, B. subtilis facilitates the careful adjustment of expression levels and genetic background required for full functionality of the introduced regulators.
Date of acceptance
2014
Autoren
  • Chong Fang
  • Emanuel Stiegeler
  • Gregory M Cook
  • Thorsten Mascher
  • Susanne Gebhard
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/24676422
DOI
10.1371/journal.pone.0093169
eISSN
1932-6203
Externe Identifier
  • PubMed Central ID: PMC3968067
Ausgabe der Veröffentlichung
3
Zeitschrift
PLoS One
Schlüsselwörter
  • Anti-Bacterial Agents
  • Bacillus subtilis
  • Cell Wall
  • Drug Resistance, Bacterial
  • Enterococcus faecalis
  • Gene Expression
  • Genes, Bacterial
  • Genes, Reporter
  • Microbial Sensitivity Tests
  • Promoter Regions, Genetic
  • Transduction, Genetic
Sprache
eng
Country
United States
Paginierung
e93169
PII
PONE-D-13-47963
Datum der Veröffentlichung
2014
Status
Published online
Datum, an dem der Datensatz öffentlich gemacht wurde
2015
Titel
Bacillus subtilis as a platform for molecular characterisation of regulatory mechanisms of Enterococcus faecalis resistance against cell wall antibiotics.
Sub types
  • Journal Article
  • Research Support, Non-U.S. Gov't
Ausgabe der Zeitschrift
9

Datenquelle: PubMed

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