The dual transcriptional regulator CysR in Corynebacterium glutamicum ATCC 13032 controls a subset of genes of the McbR regulon in response to the availability of sulphide acceptor molecules.
BACKGROUND
Regulation of sulphur metabolism in Corynebacterium glutamicum ATCC 13032 has been studied intensively in the last few years, due to its industrial as well as scientific importance. Previously, the gene cg0156 was shown to belong to the regulon of McbR, a global transcriptional repressor of sulphur metabolism in C. glutamicum. This gene encodes a putative ROK-type regulator, a paralogue of the activator of sulphonate utilisation, SsuR. Therefore, it is an interesting candidate for study to further the understanding of the regulation of sulphur metabolism in C. glutamicum.


RESULTS
Deletion of cg0156, now designated cysR, results in the inability of the mutant to utilise sulphate and aliphatic sulphonates. DNA microarray hybridisations revealed 49 genes with significantly increased and 48 with decreased transcript levels in presence of the native CysR compared to a cysR deletion mutant. Among the genes positively controlled by CysR were the gene cluster involved in sulphate reduction, fpr2 cysIXHDNYZ, and ssuR. Gel retardation experiments demonstrated that binding of CysR to DNA depends in vitro on the presence of either O-acetyl-L-serine or O-acetyl-L-homoserine. Mapping of the transcription start points of five transcription units helped to identify a 10 bp inverted repeat as the possible CysR binding site. Subsequent in vivo tests proved this motif to be necessary for CysR-dependent transcriptional regulation.


CONCLUSIONS
CysR acts as the functional analogue of the unrelated LysR-type regulator CysB from Escherichia coli, controlling sulphide production in response to acceptor availability. In both bacteria, gene duplication events seem to have taken place which resulted in the evolution of dedicated regulators for the control of sulphonate utilisation. The striking convergent evolution of network topology indicates the strong selective pressure to control the metabolism of the essential but often toxic sulphur-containing (bio-)molecules.
Institut für Systembiologie & Genomforschung, Universität Bielefeld, Universitätsstr. 25, D-33615 Bielefeld, Germany. christian.rueckert@cebitec.uni-bielefeld.de
Christian
C
Rückert
Johanna
J
Milse
Andreas
A
Albersmeier
Daniel J
DJ
Koch
Alfred
A
Pühler
Jörn
J
Kalinowski
D000476:Alkanesulfonates / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites; D048230:Corynebacterium glutamicum / Q000235:genetics* / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D005810:Multigene Family; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012329:RNA, Bacterial / Q000235:genetics; D018085:Regulon; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013431:Sulfates / Q000378:metabolism*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
D000476:Alkanesulfonates; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C483725:McbR protein, C glutamicum; D012329:RNA, Bacterial; D012097:Repressor Proteins; D013431:Sulfates; C002634:ethane sulfonate; D003545:Cysteine