Journal Article

Mutational Analysis of Regulatory <i>cis</i>-acting Elements for the Transcriptional Activation of the <i>dmsCBA</i> Operon in <i>Rhodobacter</i> <i>sphaeroides</i> f. sp. <i>denitrificans</i>

Isamu Yamamoto, Takeshi Ujiiye, Yoshinori Ohshima and Toshio Satoh

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 7, pages 703-709
Published in print July 2001 | ISSN: 0032-0781
Published online July 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce083
Mutational Analysis of Regulatory cis-acting Elements for the Transcriptional Activation of the dmsCBA Operon in Rhodobacter sphaeroides f. sp. denitrificans

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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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 dmsClacZ 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.

Keywords: Key words: cis-acting regulatory elements — Dimethyl sulfoxide reductase — dmsCBA — Promoter analysis — Response regulator DmsR — Rhodobacter sphaeroides f. sp. denitrificans.

Journal Article.  4754 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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