Direct stimulus perception and transcription activation by a membrane-bound DNA binding protein

Publication type:
Journal article
Metadata:
Autoren
Autoren-URL
https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000268792700013&DestLinkType=FullRecord&DestApp=WOS_CPL
DOI
10.1111/j.1365-2958.2009.06787.x
eISSN
1365-2958
Externe Identifier
  • Clarivate Analytics Document Solution ID: 481FL
  • PubMed Identifier: 19602149
ISSN
0950-382X
Ausgabe der Veröffentlichung
3
Zeitschrift
MOLECULAR MICROBIOLOGY
Paginierung
482 - 491
Datum der Veröffentlichung
2009
Status
Published
Titel
Direct stimulus perception and transcription activation by a membrane-bound DNA binding protein
Sub types
  • Article
Ausgabe der Zeitschrift
73

Data source: Web of Science (Lite)

Other metadata sources:
Abstract
<jats:title>Summary</jats:title><jats:p>Few membrane proteins with a role in transcriptional regulation have been studied, and none are able to perceive their respective stimuli and activate transcription of their regulons without the aid of auxiliary proteins. The bacitracin resistance regulator, BcrR, of <jats:italic>Enterococcus faecalis</jats:italic> is a membrane‐bound DNA binding protein and is required for bacitracin‐dependent expression of the bacitracin resistance genes, <jats:italic>bcrABD</jats:italic>. Here, we show that BcrR interacts directly with Zn<jats:sup>2+</jats:sup> bacitracin (<jats:italic>K</jats:italic><jats:sub>d</jats:sub> = 2–5 μM), but not metal‐free bacitracin. A solution‐based DNA binding assay demonstrated that the affinity of BcrR for its target DNA is much higher (<jats:italic>K</jats:italic><jats:sub>d</jats:sub> = 40 nM) than previously found for transmembrane regulators and is comparable to that of soluble DNA binding proteins. A construct of BcrR that lacked the transmembrane domain was unable to bind to DNA, indicating that membrane localization was important for DNA binding. Bacitracin did not cause a change in the DNaseI footprint of BcrR on the <jats:italic>bcrA</jats:italic> promoter, but <jats:italic>in vitro</jats:italic> transcription assays with BcrR proteoliposomes showed bacitracin‐dependent activation of transcription. These findings demonstrate that BcrR is a bona fide one‐component transmembrane signal transduction system, which perceives an extracellular stimulus (presence of bacitracin) and relays it to an intracellular transcriptional response independent of any auxiliary proteins.</jats:p>
Autoren
DOI
10.1111/j.1365-2958.2009.06787.x
eISSN
1365-2958
ISSN
0950-382X
Ausgabe der Veröffentlichung
3
Zeitschrift
Molecular Microbiology
Sprache
en
Online publication date
2009
Paginierung
482 - 491
Datum der Veröffentlichung
2009
Status
Published
Herausgeber
Wiley
Herausgeber URL
http://dx.doi.org/10.1111/j.1365-2958.2009.06787.x
Datum der Datenerfassung
2023
Titel
Direct stimulus perception and transcription activation by a membrane‐bound DNA binding protein
Ausgabe der Zeitschrift
73

Data source: Crossref

Abstract
Few membrane proteins with a role in transcriptional regulation have been studied, and none are able to perceive their respective stimuli and activate transcription of their regulons without the aid of auxiliary proteins. The bacitracin resistance regulator, BcrR, of Enterococcus faecalis is a membrane-bound DNA binding protein and is required for bacitracin-dependent expression of the bacitracin resistance genes, bcrABD. Here, we show that BcrR interacts directly with Zn2+ bacitracin (Kd = 2-5 micropM), but not metal-free bacitracin. A solution-based DNA binding assay demonstrated that the affinity of BcrR for its target DNA is much higher (Kd = 40 nM) than previously found for transmembrane regulators and is comparable to that of soluble DNA binding proteins. A construct of BcrR that lacked the transmembrane domain was unable to bind to DNA, indicating that membrane localization was important for DNA binding. Bacitracin did not cause a change in the DNaseI footprint of BcrR on the bcrA promoter, but in vitro transcription assays with BcrR proteoliposomes showed bacitracin-dependent activation of transcription. These findings demonstrate that BcrR is a bona fide one-component transmembrane signal transduction system, which perceives an extracellular stimulus (presence of bacitracin) and relays it to an intracellular transcriptional response independent of any auxiliary proteins.
Addresses
  • Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin, New Zealand.
Autoren
DOI
10.1111/j.1365-2958.2009.06787.x
eISSN
1365-2958
Externe Identifier
  • PubMed Identifier: 19602149
  • PubMed Central ID: PMC2752741
Funding acknowledgements
  • NIGMS NIH HHS: R01 GM047446
  • NIGMS NIH HHS: GM047446
  • NIGMS NIH HHS: R01 GM047446-13
Open access
false
ISSN
0950-382X
Ausgabe der Veröffentlichung
3
Zeitschrift
Molecular microbiology
Schlüsselwörter
  • Enterococcus faecalis
  • Zinc
  • Bacitracin
  • Bacterial Proteins
  • ATP-Binding Cassette Transporters
  • DNA-Binding Proteins
  • DNA, Bacterial
  • Signal Transduction
  • Gene Expression Regulation, Bacterial
  • Transcriptional Activation
Sprache
eng
Medium
Print-Electronic
Online publication date
2009
Paginierung
482 - 491
Datum der Veröffentlichung
2009
Status
Published
Datum der Datenerfassung
2009
Titel
Direct stimulus perception and transcription activation by a membrane-bound DNA binding protein.
Sub types
  • research-article
  • Journal Article
  • Research Support, N.I.H., Extramural
Ausgabe der Zeitschrift
73

Files

https://onlinelibrary.wiley.com/doi/pdfdirect/10.1111/j.1365-2958.2009.06787.x https://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19602149/pdf/?tool=EBI https://europepmc.org/articles/PMC2752741?pdf=render

Data source: Europe PubMed Central

Abstract
Few membrane proteins with a role in transcriptional regulation have been studied, and none are able to perceive their respective stimuli and activate transcription of their regulons without the aid of auxiliary proteins. The bacitracin resistance regulator, BcrR, of Enterococcus faecalis is a membrane-bound DNA binding protein and is required for bacitracin-dependent expression of the bacitracin resistance genes, bcrABD. Here, we show that BcrR interacts directly with Zn2+ bacitracin (Kd = 2-5 micropM), but not metal-free bacitracin. A solution-based DNA binding assay demonstrated that the affinity of BcrR for its target DNA is much higher (Kd = 40 nM) than previously found for transmembrane regulators and is comparable to that of soluble DNA binding proteins. A construct of BcrR that lacked the transmembrane domain was unable to bind to DNA, indicating that membrane localization was important for DNA binding. Bacitracin did not cause a change in the DNaseI footprint of BcrR on the bcrA promoter, but in vitro transcription assays with BcrR proteoliposomes showed bacitracin-dependent activation of transcription. These findings demonstrate that BcrR is a bona fide one-component transmembrane signal transduction system, which perceives an extracellular stimulus (presence of bacitracin) and relays it to an intracellular transcriptional response independent of any auxiliary proteins.
Autoren
Autoren-URL
https://www.ncbi.nlm.nih.gov/pubmed/19602149
DOI
10.1111/j.1365-2958.2009.06787.x
eISSN
1365-2958
Externe Identifier
  • NIH Manuscript Submission ID: NIHMS134991
  • PubMed Central ID: PMC2752741
Funding acknowledgements
  • NIGMS NIH HHS: R01 GM047446
  • NIGMS NIH HHS: R01 GM047446-13
  • NIGMS NIH HHS: GM047446
Ausgabe der Veröffentlichung
3
Zeitschrift
Mol Microbiol
Schlüsselwörter
  • ATP-Binding Cassette Transporters
  • Bacitracin
  • Bacterial Proteins
  • DNA, Bacterial
  • DNA-Binding Proteins
  • Enterococcus faecalis
  • Gene Expression Regulation, Bacterial
  • Signal Transduction
  • Transcriptional Activation
  • Zinc
Sprache
eng
Country
England
Paginierung
482 - 491
PII
MMI6787
Datum der Veröffentlichung
2009
Status
Published
Datum, an dem der Datensatz öffentlich gemacht wurde
2009
Titel
Direct stimulus perception and transcription activation by a membrane-bound DNA binding protein.
Sub types
  • Journal Article
  • Research Support, N.I.H., Extramural
Ausgabe der Zeitschrift
73

Data source: PubMed

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