title: Genome-wide analysis of the role of global transcriptional regulator GntR1 in Corynebacterium glutamicum.
abstract: 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.
authors: Research Institute of Innovative Technology for the Earth, Kyoto, Japan.
authors: Yuya
authors: Y
authors: Tanaka
authors: Research Institute of Innovative Technology for the Earth, Kyoto, Japan.
authors: Norihiko
authors: N
authors: Takemoto
authors: Research Institute of Innovative Technology for the Earth, Kyoto, Japan Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara, Japan.
authors: Terukazu
authors: T
authors: Ito
authors: Research Institute of Innovative Technology for the Earth, Kyoto, Japan.
authors: Haruhiko
authors: H
authors: Teramoto
authors: Research Institute of Innovative Technology for the Earth, Kyoto, Japan.
authors: Hideaki
authors: H
authors: Yukawa
authors: Research Institute of Innovative Technology for the Earth, Kyoto, Japan Graduate School of Biological Sciences, Nara Institute of Science and Technology, Nara, Japan mmg-lab@rite.or.jp.
authors: Masayuki
authors: M
authors: Inui
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D048230:Corynebacterium glutamicum / Q000235:genetics / Q000378:metabolism*; D004269:DNA, Bacterial; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology*; D016680:Genome, Bacterial; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D050436:Regulatory Elements, Transcriptional; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial