|Genome-wide analysis of the role of global transcriptional regulator GntR1
in Corynebacterium glutamicum.
J. Bacteriol. 196: 3249-3258. 2014.
Y. Tanaka, N. Takemoto, T. Ito, H. Teramoto, H. Yukawa and M. Inui.
|The transcriptional regulator GntR1 downregulates the genes for gluconate
catabolism and pentose phosphate pathway in Corynebacterium glutamicum. Gluconate lowers the DNA binding affinity of GntR1, which is probably the mechanism of gluconate-dependent induction of these genes. In addition, GntR1 positively regulates ptsG, a gene encoding a major glucose transporter, and pck, a gene encoding phosphoenolpyruvate carboxykinase. Here, we searched for the new target of GntR1 on a genome-wide scale by chromatin immunoprecipitation in conjunction with microarray (ChIP-chip) analysis. This analysis identified 56 in vivo GntR1 binding sites, of which 7 sites were previously reported. The newly
identified GntR1 sites include the upstream regions of carbon metabolism
genes such as pyk, maeB, gapB, and icd, encoding pyruvate kinase, malic enzyme, glyceraldehyde 3-phosphate dehydrogenase
B, and isocitrate dehydrogenase, respectively. Binding of GntR1 to the
promoter region of these genes was confirmed by electrophoretic mobility
shift assay. The activity of the icd, gapB, and maeB promoters was reduced by the mutation at the GntR1 binding site, in contrast
to the pyk promoter activity, which was increased, indicating that GntR1 is a transcriptional activator of icd, gapB, and maeB and is a repressor of pyk. Thus, it is likely that GntR1 stimulates glucose uptake by inducing the
phosphoenolpyruvate (PEP):carbohydrate phosphotransferase system (PTS)
gene while repressing pyk to increase PEP availability in the absence of gluconate. Repression of
zwf and gnd may reduce the NADPH supply, which may be compensated by the induction
of maeB and icd. Upregulation of icd, gapB, and maeB and downregulation of pyk by GntR1 probably support gluconeogenesis.