Direct stimulus perception and transcription activation by a membrane-bound DNA binding protein.
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.
Gebhard
Susanne
S
Department of Microbiology and Immunology, University of Otago, PO Box 56, Dunedin, New Zealand.
Gaballa
Ahmed
A
Helmann
John D
JD
Cook
Gregory M
GM
D018528:ATP-Binding Cassette Transporters / Q000235:genetics / Q000378:metabolism*; D001414:Bacitracin / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004269:DNA, Bacterial / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D013293:Enterococcus faecalis / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015398:Signal Transduction; D015533:Transcriptional Activation; D015032:Zinc / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; C493252:BcrR protein, Enterococcus faecalis; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D001414:Bacitracin; D015032:Zinc