Mutational analysis of regulatory cis-acting elements for the transcriptional activation of the dmsCBA operon in Rhodobacter sphaeroides f. sp. denitrificans.
Four direct repeats of a 10-nt sequence, called dms boxes, are located upstream of the dmsCBA operon encoding dimethyl sulfoxide (DMSO) reductase in Rhodobacter sphaeroides f. sp. denitrificans IL106. Two dms boxes 1 and 2 have been shown to be binding sites of DmsR protein, a response regulator of a two-component system involved in the anaerobic induction by DMSO of DMSO reductase synthesis. In this study, functions of four dms boxes in the transcriptional regulation of the dmsCBA operon were investigated. The transcription start site of the dmsCBA genes was identified at the distance of 23 nt downstream of the closest dms box 4. Expression of the dmsC-lacZ gene fusion which included the dmsCBA promoter region containing the dms boxes was examined and its anaerobic induction by DMSO and DmsR-dependency were demonstrated in the phototroph. The examination with nucleotide substitutions in the four respective dms boxes showed that the set of four dms boxes is required for the dmsCBA operon activation. Moreover, the importance of the nucleotide sequence of TTCAC in dms box 4 and of A at the center in dms box 1 was significantly shown. These facts suggest that the pentad nucleotides TTCAC and TTAAC in the dms boxes serve as cis-acting elements in the transcriptional activation of the dmsCBA operon.
Yamamoto
I
I
Department of Biological Science, Graduate School of Science, Hiroshima University, Higashi-Hiroshima, 739-8526, Japan. iyamamo@hiroshima-u.ac.jp
Ujiiye
T
T
Ohshima
Y
Y
Satoh
T
T
D000693:Anaerobiosis; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins; D001483:Base Sequence; D003580:Cytochromes / Q000235:genetics* / Q000378:metabolism; D004252:DNA Mutational Analysis / Q000379:methods; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins; D005809:Genes, Regulator; D007506:Iron-Sulfur Proteins; D007763:Lac Operon / Q000235:genetics; D008969:Molecular Sequence Data; D009876:Operon; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012242:Rhodobacter sphaeroides / Q000201:enzymology / Q000235:genetics* / Q000378:metabolism; D015533:Transcriptional Activation
D001426:Bacterial Proteins; D003580:Cytochromes; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; C102556:DmsC protein, E coli; C107311:DmsR protein, Rhodobacter sphaeroides; D029968:Escherichia coli Proteins; D007506:Iron-Sulfur Proteins; D010088:Oxidoreductases; C045664:dimethyl sulfoxide reductase