Rückert, Christian; Milse, Johanna; Albersmeier, Andreas; Koch, Daniel J.; Pühler, Alfred; Kalinowski, Jörn: The dual transcriptional regulator CysR in Corynebacterium glutamicum ATCC 13032 controls a subset of genes of the McbR regulon in response to the availability [...]. In: BMC GENOMICS. Jg.9 H. 1. 2008, S. 483 ff.
Inhalt
- 1 Background
- 2 Results
- 2.1 The Corynebacterium glutamicum mutant CR030 with a deleted cg0156 (cysR) gene can no longer grow with sulphate or sulphonates as sole source of sulphur
- 2.2 The regulator CysR affects a subset of genes of the McbR regulon
- 2.3 O-acetyl-L-serine (OAS) and O-acetyl-L-homoserine (OAH) are required as effector substances for the binding of the regulator CysR to DNA
- 2.4 A 10 bp inverted repeat is present in the mapped promoter/operator region of CysR-controlled genes
- 2.5 CysR-dependent regulation requires the presence of the identified 10 bp inverted repeat in vivo
- 2.6 The proposed regulon of the dual regulator CysR consists of seven transcription units
- 3 Discussion
- 3.1 The CysR protein is involved in the activation of the pathway for assimilatory sulphate reduction and possibly related genes in C. glutamicum
- 3.2 The regulator CysR is a member of the ROK protein family
- 3.3 The cysR gene is transcribed as leaderless transcript
- 3.4 The presence of sulphide acceptor molecules is necessary for binding of CysR to DNA
- 3.5 The deduced regulatory model of CysR in C. glutamicum
- Conclusion
- 4 Methods
- 4.1 Bacterial strains, plasmids and culture media
- 4.2 DNA isolation, transfer and manipulation
- 4.3 Polymerase chain reaction experiments
- 4.4 Construction of plasmids
- 4.5 Site-specific gene deletion/promoter replacement
- 4.6 Real-time monitoring of cell growth using nephelometry
- 4.7 RNA preparation and DNA microarray hybridisation
- 4.8 Purification of the CysR protein
- 4.9 Electrophoretic mobility shift assays
- 4.10 Determination of transcriptional starts with the RACE method
- 4.11 Relative mRNA quantification using real-time RT- PCR
- 4.12 GenBank/TrEMBL accession numbers
- 4.13 Bioinformatic analysis
- Authors' contributions
- Acknowledgements
- References