Protein binding in vivo to OP2 promoter of the Pseudomonas putida TOL plasmid.
The transcription of OP2 encoding enzymes for m-toluate catabolism on the Pseudomonas putida TOL plasmid is activated by basal-level XylS protein in the presence of m-toluate or by overproduced XylS protein in the absence of m-toluate. In this study, in vivo dimethyl sulfate (DMS) footprinting was performed to understand the mechanism of transcriptional regulation of OP2 promoter by XylS. In the presence of overproduced XylS without m-toluate, several protected nucleotides were observed, indicating the binding of RNA polymerase to DNA. However, the protection was canceled upon addition of m-toluate. These results suggest that RNA polymerase is retained by XylS on the OP2 promoter in the absence of inducer, and is released by m-toluate binding to XylS, concomitant with transcription.
Miura
K
K
Department of Biochemistry, Yamaguchi University School of Medicine, Japan.
Inouye
S
S
Nakazawa
A
A
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001565:Benzoates / Q000378:metabolism / Q000494:pharmacology; D018983:DNA Footprinting; D019175:DNA Methylation / Q000187:drug effects; D004268:DNA-Binding Proteins / Q000378:metabolism*; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005798:Genes, Bacterial / Q000235:genetics*; D009876:Operon; D010957:Plasmids / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding; D016958:Pseudomonas putida / Q000235:genetics*; D012293:Rifampin / Q000494:pharmacology; D013463:Sulfuric Acid Esters / Q000494:pharmacology; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects
D001426:Bacterial Proteins; D001565:Benzoates; D004268:DNA-Binding Proteins; D013463:Sulfuric Acid Esters; D015534:Trans-Activators; C078720:XylS protein, Pseudomonas putida; C009503:3-toluic acid; D012321:DNA-Directed RNA Polymerases; C007482:dimethyl sulfate; D012293:Rifampin