Transcriptional control of the succinate dehydrogenase operon sdhCAB of Corynebacterium glutamicum by the cAMP-dependent regulator GlxR and the LuxR-type regulator RamA.
In experiments performed to identify transcriptional regulators of the tricarboxylic acid cycle of Corynebacterium glutamicum, the cAMP-dependent regulator GlxR and the regulators of acetate metabolism RamA and RamB were enriched by DNA affinity chromatography with the promoter region of the sdhCAB operon encoding succinate dehydrogenase. The binding of purified GlxR, RamA and RamB was verified by electrophoretic mobility shift assays and the regulatory effects of these proteins on sdhCAB gene expression were tested by promoter activity assays and SDH activity measurements. Evidence was obtained that GlxR functions as a repressor and RamA as an activator of sdhCAB expression, whereas RamB had no obvious influence under the conditions tested.
Bussmann
Michael
M
Institut für Biotechnologie 1, Forschungszentrum Jülich, D-52425 Jülich, Germany.
Emer
Denise
D
Hasenbein
Sonja
S
Degraf
Sabine
S
Eikmanns
Bernhard J
BJ
Bott
Michael
M
D001426:Bacterial Proteins / Q000302:isolation & purification / Q000378:metabolism*; D001483:Base Sequence; D002846:Chromatography, Affinity; D048230:Corynebacterium glutamicum / Q000201:enzymology* / Q000235:genetics; D000242:Cyclic AMP / Q000378:metabolism*; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D021901:DNA, Intergenic / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D009876:Operon / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011485:Protein Binding; D012097:Repressor Proteins / Q000378:metabolism*; D013385:Succinate Dehydrogenase / Q000235:genetics*; D015534:Trans-Activators / Q000378:metabolism*; D024363:Transcription Initiation Site; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D017931:DNA Primers; D004269:DNA, Bacterial; D021901:DNA, Intergenic; D012097:Repressor Proteins; D015534:Trans-Activators; C064778:LuxR autoinducer binding proteins; D000242:Cyclic AMP; D013385:Succinate Dehydrogenase