Functional characterization of BcrR: a one-component transmembrane signal transduction system for bacitracin resistance
- Publication type:
- Journal article
- Metadata:
-
- Autoren
- Rachel L Darnell
- Yoshio Nakatani
- Melanie K Knottenbelt
- Susanne Gebhard
- Gregory M Cook
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000466843400011&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1099/mic.0.000781
- eISSN
- 1465-2080
- Externe Identifier
- Clarivate Analytics Document Solution ID: HW7AY
- PubMed Identifier: 30777814
- ISSN
- 1350-0872
- Ausgabe der Veröffentlichung
- 4
- Zeitschrift
- MICROBIOLOGY-SGM
- Schlüsselwörter
- Enterococcus
- antimicrobial resistance
- membrane protein
- regulator
- Paginierung
- 475 - 487
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Titel
- Functional characterization of BcrR: a one-component transmembrane signal transduction system for bacitracin resistance
- Sub types
- Article
- Ausgabe der Zeitschrift
- 165
Data source: Web of Science (Lite)
- Other metadata sources:
-
- Autoren
- Rachel L Darnell
- Yoshio Nakatani
- Melanie K Knottenbelt
- Susanne Gebhard
- Gregory M Cook
- DOI
- 10.1099/mic.0.000781
- eISSN
- 1465-2080
- ISSN
- 1350-0872
- Ausgabe der Veröffentlichung
- 4
- Zeitschrift
- Microbiology
- Sprache
- en
- Paginierung
- 475 - 487
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Herausgeber
- Microbiology Society
- Herausgeber URL
- http://dx.doi.org/10.1099/mic.0.000781
- Datum der Datenerfassung
- 2019
- Titel
- Functional characterization of BcrR: a one-component transmembrane signal transduction system for bacitracin resistance
- Ausgabe der Zeitschrift
- 165
Data source: Crossref
- Abstract
- Bacitracin is a cell wall targeting antimicrobial with clinical and agricultural applications. With the growing mismatch between antimicrobial resistance and development, it is essential we understand the molecular mechanisms of resistance in order to prioritize and generate new effective antimicrobials. BcrR is a unique membrane-bound one-component system that regulates high-level bacitracin resistance in Enterococcus faecalis. In the presence of bacitracin, BcrR activates transcription of the bcrABD operon conferring resistance through a putative ATP-binding cassette (ABC) transporter (BcrAB). BcrR has three putative functional domains, an N-terminal helix-turn-helix DNA-binding domain, an intermediate oligomerization domain and a C-terminal transmembrane domain. However, the molecular mechanisms of signal transduction remain unknown. Random mutagenesis of bcrR was performed to generate loss- and gain-of-function mutants using transcriptional reporters fused to the target promoter PbcrA. Fifteen unique mutants were isolated across all three proposed functional domains, comprising 14 loss-of-function and one gain-of-function mutant. The gain-of-function variant (G64D) mapped to the putative dimerization domain of BcrR, and functional analyses indicated that the G64D mutant constitutively expresses the PbcrA-luxABCDE reporter. DNA-binding and membrane insertion were not affected in the five mutants chosen for further characterization. Homology modelling revealed putative roles for two key residues (R11 and S33) in BcrR activation. Here we present a new model of BcrR activation and signal transduction, providing valuable insight into the functional characterization of membrane-bound one-component systems and how they can coordinate critical bacterial responses, such as antimicrobial resistance.
- Addresses
- 1Department of Microbiology and Immunology, University of Otago, Dunedin, New zealand.
- Autoren
- Rachel L Darnell
- Yoshio Nakatani
- Melanie K Knottenbelt
- Susanne Gebhard
- Gregory M Cook
- DOI
- 10.1099/mic.0.000781
- eISSN
- 1465-2080
- Externe Identifier
- PubMed Identifier: 30777814
- Open access
- false
- ISSN
- 1350-0872
- Ausgabe der Veröffentlichung
- 4
- Zeitschrift
- Microbiology (Reading, England)
- Schlüsselwörter
- Cell Membrane
- Bacillus subtilis
- Enterococcus faecalis
- Bacitracin
- ATP-Binding Cassette Transporters
- Anti-Bacterial Agents
- Drug Resistance, Bacterial
- Signal Transduction
- Gene Expression Regulation, Bacterial
- Protein Binding
- Mutation
- Genes, Bacterial
- Operon
- Promoter Regions, Genetic
- Protein Domains
- Sprache
- eng
- Medium
- Print-Electronic
- Online publication date
- 2019
- Paginierung
- 475 - 487
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Datum der Datenerfassung
- 2019
- Titel
- Functional characterization of BcrR: a one-component transmembrane signal transduction system for bacitracin resistance.
- Sub types
- Research Support, Non-U.S. Gov't
- Journal Article
- Ausgabe der Zeitschrift
- 165
Data source: Europe PubMed Central
- Abstract
- Bacitracin is a cell wall targeting antimicrobial with clinical and agricultural applications. With the growing mismatch between antimicrobial resistance and development, it is essential we understand the molecular mechanisms of resistance in order to prioritize and generate new effective antimicrobials. BcrR is a unique membrane-bound one-component system that regulates high-level bacitracin resistance in Enterococcus faecalis. In the presence of bacitracin, BcrR activates transcription of the bcrABD operon conferring resistance through a putative ATP-binding cassette (ABC) transporter (BcrAB). BcrR has three putative functional domains, an N-terminal helix-turn-helix DNA-binding domain, an intermediate oligomerization domain and a C-terminal transmembrane domain. However, the molecular mechanisms of signal transduction remain unknown. Random mutagenesis of bcrR was performed to generate loss- and gain-of-function mutants using transcriptional reporters fused to the target promoter PbcrA. Fifteen unique mutants were isolated across all three proposed functional domains, comprising 14 loss-of-function and one gain-of-function mutant. The gain-of-function variant (G64D) mapped to the putative dimerization domain of BcrR, and functional analyses indicated that the G64D mutant constitutively expresses the PbcrA-luxABCDE reporter. DNA-binding and membrane insertion were not affected in the five mutants chosen for further characterization. Homology modelling revealed putative roles for two key residues (R11 and S33) in BcrR activation. Here we present a new model of BcrR activation and signal transduction, providing valuable insight into the functional characterization of membrane-bound one-component systems and how they can coordinate critical bacterial responses, such as antimicrobial resistance.
- Autoren
- Rachel L Darnell
- Yoshio Nakatani
- Melanie K Knottenbelt
- Susanne Gebhard
- Gregory M Cook
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/30777814
- DOI
- 10.1099/mic.0.000781
- eISSN
- 1465-2080
- Ausgabe der Veröffentlichung
- 4
- Zeitschrift
- Microbiology (Reading)
- Schlüsselwörter
- Enterococcus
- antimicrobial resistance
- membrane protein
- regulator
- ATP-Binding Cassette Transporters
- Anti-Bacterial Agents
- Bacillus subtilis
- Bacitracin
- Cell Membrane
- Drug Resistance, Bacterial
- Enterococcus faecalis
- Gene Expression Regulation, Bacterial
- Genes, Bacterial
- Mutation
- Operon
- Promoter Regions, Genetic
- Protein Binding
- Protein Domains
- Signal Transduction
- Sprache
- eng
- Country
- England
- Paginierung
- 475 - 487
- Datum der Veröffentlichung
- 2019
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2020
- Titel
- Functional characterization of BcrR: a one-component transmembrane signal transduction system for bacitracin resistance.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 165
Data source: PubMed
- Beziehungen:
- Property of