A protein that activates expression of a multidrug efflux transporter upon binding the transporter substrates.
Multidrug transporters are membrane proteins which, by an unknown mechanism, recognize diverse toxic compounds and efflux them from cells. We found that two substrates of the Bacillus subtilis multidrug transporter Bmr, rhodamine 6G and tetraphenylphosphonium (TPP), enhance Bmr expression at the level of transcription. Gene knock-out experiments demonstrated that an open reading frame located immediately downstream of the bmr gene is required for this enhancement. The protein product of this open reading frame, BmrR, shows distinct sequence homology to several known bacterial transcription activator proteins, such as MerR and TipAL. Gel-mobility shift and DNase protection assays indicated that BmrR binds specifically, as a dimer, to the bmr gene promoter. Furthermore, the affinity of this binding was enhanced by rhodamine and TPP, thus suggesting that these structurally dissimilar molecules interact directly with BmrR. Indeed, we found that BmrR bound rhodamine 6G stoichiometrically, one rhodamine molecule/BmrR dimer, and that TPP competed with rhodamine for this binding. Our results indicate that the enhancement of Bmr expression by some of its substrates is due to the ability of the regulatory protein, BmrR, to bind structurally dissimilar compounds resulting in enhanced transcription of the transporter gene.
Department of Medicinal Chemistry and Pharmacognosy, University of Illinois, Chicago 60612.
M
M
Ahmed
C M
CM
Borsch
S S
SS
Taylor
N
N
Vázquez-Laslop
A A
AA
Neyfakh
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000096:biosynthesis*; D000595:Amino Acid Sequence; D000970:Antineoplastic Agents / Q000494:pharmacology*; D001412:Bacillus subtilis / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D015152:Blotting, Northern; D002352:Carrier Proteins; D017931:DNA Primers / Q000378:metabolism; D004790:Enzyme Induction / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects*; D005798:Genes, Bacterial; D007700:Kinetics; D026901:Membrane Transport Proteins; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D009861:Onium Compounds / Q000494:pharmacology*; D009943:Organophosphorus Compounds / Q000494:pharmacology*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012235:Rhodamines / Q000494:pharmacology*; D017386:Sequence Homology, Amino Acid; D015534:Trans-Activators / Q000737:chemistry / Q000378:metabolism; D001616:beta-Galactosidase / Q000096:biosynthesis
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000970:Antineoplastic Agents; D001426:Bacterial Proteins; C098722:BmrR protein, bacteria; D002352:Carrier Proteins; D017931:DNA Primers; D026901:Membrane Transport Proteins; D009861:Onium Compounds; D009943:Organophosphorus Compounds; D012235:Rhodamines; D015534:Trans-Activators; C026188:rhodamine 6G; C075041:Bmr protein, Bacillus subtilis; D001616:beta-Galactosidase; C013289:tetraphenylphosphonium