The transcriptional regulator SsuR activates expression of theCorynebacterium glutamicumsulphonate utilization genes in the absence of sulphate
- 19 September 2005
- journal article
- Published by Wiley in Molecular Microbiology
- Vol. 58 (2) , 480-494
- https://doi.org/10.1111/j.1365-2958.2005.04836.x
Abstract
In a recent study, the putative regulatory gene cg0012 was shown to belong to the regulon of McbR, a global transcriptional regulator of sulphur metabolism in Corynebacterium glutamicum ATCC 13032. A deletion of cg0012, now designated ssuR (sulphonate sulphur utilization regulator), led to the mutant strain C. glutamicum DK100, which was shown to be blocked in the utilization of sulphonates as sulphur sources. According to DNA microarray hybridizations, transcription of the ssu and seu genes, encoding the sulphonate utilization system of C. glutamicum, was considerably decreased in C. glutamicum DK100 when compared with the wild-type strain. Electrophoretic mobility shift assays with purified SsuR protein demonstrated that the upstream regions of ssuI, seuABC, ssuD2 and ssuD1CBA contain SsuR binding sites. A nucleotide sequence alignment of the four DNA fragments containing the SsuR binding sites revealed a common 21 bp motif consisting of T-, GC- and A-rich domains. Mapping of the transcriptional start sites in front of ssuI, seuABC, ssuD2 and ssuD1CBA indicated that the SsuR binding sites are located directly upstream of identified promoter sequences and that the ssu genes are expressed by leaderless transcripts. Binding of the SsuR protein to its operator was shown to be diminished in vitro by the effector substance sulphate and its direct assimilation products adenosine 5'-phosphosulphate, sulphite and sulphide. Real-time reverse transcription polymerase chain reaction experiments verified that the expression of the ssu and seu genes was also repressed in vivo by the presence of sulphate or sulphite. Therefore, the regulatory protein SsuR activates the expression of the ssu and seu genes in C. glutamicum in the absence of the preferred sulphur source sulphate.Keywords
This publication has 40 references indexed in Scilit:
- Role of thessuandseuGenes ofCorynebacterium glutamicumATCC 13032 in Utilization of Sulfonates and Sulfonate Esters as Sulfur SourcesApplied and Environmental Microbiology, 2005
- The McbR repressor modulated by the effector substanceS‐adenosylhomocysteine controls directly the transcription of a regulon involved in sulphur metabolism ofCorynebacterium glutamicumATCC 13032Molecular Microbiology, 2005
- Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extensionPublished by Elsevier ,2003
- Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicumPublished by Elsevier ,2003
- Efficient Electrotransformation of Corynebacterium diphtheriae with a Mini-Replicon Derived from the Corynebacterium glutamicum Plasmid pGA1Current Microbiology, 2002
- Functional analysis of the Bacillus subtilis cysK and cysJI genesFEMS Microbiology Letters, 2001
- Probability-based protein identification by searching sequence databases using mass spectrometry dataElectrophoresis, 1999
- Regulation of the sulfate starvation response in Pseudomonas aeruginosa: role of cysteine biosynthetic intermediatesMicrobiology, 1998
- Evolutionary relationships between sugar kinases and transcriptional repressors in bacteriaMicrobiology, 1994
- Cloning and characterization of the N-acetylglucosamine operon of Escherichia coliBiochemistry and Cell Biology, 1990