Gaigalat, Lars; Schlüter, Jan-Philip; Hartmann, Michelle; Mormann, Sascha; Tauch, Andreas; Pühler, Alfred ; Kalinowski, Jörn: The DeoR-type transcriptional regulator SugR acts as a repressor for genes encoding the phosphoenolpyruvate: sugar phosphotransferase system (PTS) in Corynebacterium [...]. In: BMC Molecular Biology. Jg.8 H. 1. 2007
Inhalt
- Conclusion
- Background
- Results
- The extended fructose-PTS gene cluster of C. glutamicum contains four transcriptional units
- Transcription of the fructose-PTS gene cluster is enhanced in C. glutamicum cultures grown on glucose or fructose when compared to cultures grown on acetate
- The sugR gene is involved in the transcriptional regulation of the fructose-PTS cluster containing ptsI, cg2118, fruK, ptsF, and ptsH of C. glutamicum
- The sugR gene of C. glutamicum is involved in the repression of PTS gene transcription
- The SugR protein of C. glutamicum binds to sequences located upstream of the coding regions of ptsI, cg2118, ptsH, ptsG, and ptsS
- The C. glutamicum SugR repressor binds to two 21 bp DNA regions interfering with ptsI and cg2118-fruK-ptsF transcription
- The effectors fructose-1-phosphate, fructose-1,6- bisphosphate, and glucose-6-phosphate influence the binding activity of the SugR repressor
- Discussion
- The C. glutamicum regulatory gene sugR is involved in the repression of the PTS genes
- The DeoR-type regulator SugR acts as a repressor on transcription of all PTS genes in C. glutamicum
- The SugR binding site comprises a 21-base pair AC-rich motif
- Fructose-1-phosphate and other sugar phosphates act as effectors releasing SugR from its binding sites
- Conclusion
- Methods
- Strains and media
- PCR techniques
- DNA isolation, transfer and manipulation
- Construction of the C. glutamicum deletion mutant strains
- Genetic construction, expression and purification of heterologous expressed Intein-coupled protein SugR
- Separation of cytoplasmic proteins of C. glutamicum RES167
- Operator binding assays by electrophoretic mobility shift assay (EMSA) with purified SugR
- Total RNA Isolation from C. glutamicum cultures
- Determination of transcription starts by 5'-RACE
- Relative quantification of mRNA levels using real-time RT- PCR
- Microarray experiments and analysis
- Bioinformatic tools used to analyse nucleotide sequences
- Authors' contributions
- Acknowledgements
- References