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      Identification of the Regulators Binding to the Upstream Region of glxR in Corynebacterium glutamicum = Identification of the Regulators Binding to the Upstream Region of glxR in Corynebacterium glutamicum

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      https://www.riss.kr/link?id=A100767915

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      다국어 초록 (Multilingual Abstract)

      GlxR is considered as a global transcriptional regulator controlling a large number of genes having broad physiological aspects in Corynebacterium glutamicum. However, the expression profile revealing the transcriptional control of glxR has not yet been studied in detail. DNA affinity chromatography experiments revealed the binding of transcriptional regulators SucR, RamB, GlxR, and a GntR-type protein (hereafter denoted as GntR3) to the upstream region of glxR. The binding of different regulators to the glxR promoter was confirmed by EMSA experiments. The expression of glxR was analyzed in detail under various carbon sources in the wild-type and different mutant strains. The sucR and gntR3 deletion mutants showed decreased glxR promoter activities, when compared with the wild type, irrespective of the carbon sources. The promoter activity of glxR was derepressed in the ramB deletion mutant under all the tested carbon sources. These results indicate that SucR and GntR3 are acting as activators of GlxR, while RamB plays a repressor. As expected, the expression of glxR in the cyaB and glxR deletion mutants was derepressed under different media conditions, indicating that GlxR is autoregulated.
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      GlxR is considered as a global transcriptional regulator controlling a large number of genes having broad physiological aspects in Corynebacterium glutamicum. However, the expression profile revealing the transcriptional control of glxR has not yet be...

      GlxR is considered as a global transcriptional regulator controlling a large number of genes having broad physiological aspects in Corynebacterium glutamicum. However, the expression profile revealing the transcriptional control of glxR has not yet been studied in detail. DNA affinity chromatography experiments revealed the binding of transcriptional regulators SucR, RamB, GlxR, and a GntR-type protein (hereafter denoted as GntR3) to the upstream region of glxR. The binding of different regulators to the glxR promoter was confirmed by EMSA experiments. The expression of glxR was analyzed in detail under various carbon sources in the wild-type and different mutant strains. The sucR and gntR3 deletion mutants showed decreased glxR promoter activities, when compared with the wild type, irrespective of the carbon sources. The promoter activity of glxR was derepressed in the ramB deletion mutant under all the tested carbon sources. These results indicate that SucR and GntR3 are acting as activators of GlxR, while RamB plays a repressor. As expected, the expression of glxR in the cyaB and glxR deletion mutants was derepressed under different media conditions, indicating that GlxR is autoregulated.

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      참고문헌 (Reference)

      1 Studier FW, "Use of bacteriophage T7RNA polymerase to direct selective high-level expression of cloned genes" 189 : 113-130, 1986

      2 Jungwirth B, "Triple transcriptional control of the resuscitation promoting factor 2 (rpf2) gene of Corynebacterium glutamicum by the regulators of acetate metabolism RamA and RamB and the cAMP-dependent regulator GlxR" 281 : 190-197, 2008

      3 Kotrbova-Kozak A, "Transcriptionally regulated adhA gene encodes alcohol dehydrogenase required for ethanol and n-propanol utilization in Corynebacterium glutamicum R" 76 : 1347-1356, 2007

      4 Bussmann M, "Transcriptional control of the succinate dehydrogenase operon sdhCAB of Corynebacterium glutamicum by the cAMP-dependent regulator GlxR and the LuxR-type regulator RamA." 143 : 173-182, 2009

      5 Fujita Y, "The gluconate operon gnt of Bacillus subtilis encodes its own transcriptional negative regulator" 84 : 4524-4528, 1987

      6 Arndt A, "The alcohol dehydrogenase gene adhA in Corynebacterium glutamicum is subject to carbon catabolite repression" 189 : 7408-7416, 2007

      7 Kohl TA, "The GlxR regulon of the amino acid producer Corynebacterium glutamicum: in silico and in vitro detection of DNA binding sites of a global transcription regulator" 135 : 340-350, 2008

      8 Kohl TA, "The GlxR regulon of the amino acid producer Corynebacterium glutamicum: detection of the corynebacterial core regulon and integration into the transcriptional regulatory network model" 143 : 239-246, 2009

      9 Engels V, "The DeoR-type regulator SugR represses expression of ptsG in Corynebacterium glutamicum" 189 : 2955-2966, 2007

      10 Ikeda M, "The Corynebacterium glutamicum genome: features and impacts on biotechnological processes" 62 : 99-109, 2003

      1 Studier FW, "Use of bacteriophage T7RNA polymerase to direct selective high-level expression of cloned genes" 189 : 113-130, 1986

      2 Jungwirth B, "Triple transcriptional control of the resuscitation promoting factor 2 (rpf2) gene of Corynebacterium glutamicum by the regulators of acetate metabolism RamA and RamB and the cAMP-dependent regulator GlxR" 281 : 190-197, 2008

      3 Kotrbova-Kozak A, "Transcriptionally regulated adhA gene encodes alcohol dehydrogenase required for ethanol and n-propanol utilization in Corynebacterium glutamicum R" 76 : 1347-1356, 2007

      4 Bussmann M, "Transcriptional control of the succinate dehydrogenase operon sdhCAB of Corynebacterium glutamicum by the cAMP-dependent regulator GlxR and the LuxR-type regulator RamA." 143 : 173-182, 2009

      5 Fujita Y, "The gluconate operon gnt of Bacillus subtilis encodes its own transcriptional negative regulator" 84 : 4524-4528, 1987

      6 Arndt A, "The alcohol dehydrogenase gene adhA in Corynebacterium glutamicum is subject to carbon catabolite repression" 189 : 7408-7416, 2007

      7 Kohl TA, "The GlxR regulon of the amino acid producer Corynebacterium glutamicum: in silico and in vitro detection of DNA binding sites of a global transcription regulator" 135 : 340-350, 2008

      8 Kohl TA, "The GlxR regulon of the amino acid producer Corynebacterium glutamicum: detection of the corynebacterial core regulon and integration into the transcriptional regulatory network model" 143 : 239-246, 2009

      9 Engels V, "The DeoR-type regulator SugR represses expression of ptsG in Corynebacterium glutamicum" 189 : 2955-2966, 2007

      10 Ikeda M, "The Corynebacterium glutamicum genome: features and impacts on biotechnological processes" 62 : 99-109, 2003

      11 Hanahan D., "Studies on transformation of Escherichia coli with plasmids" 166 : 557-580, 1983

      12 Kronemeyer W, "Structure of the g luA BCD cluster encoding the glutamate uptake system of Corynebacterium glutamicum" 177 : 1152-1158, 1995

      13 Schäfer A, "Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum" 145 : 69-73, 1994

      14 Gerstmeir R, "RamB, a novel transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum" 186 : 2798-2809, 2004

      15 Blombach B, "RamB is an activator of the pyruvate dehydrogenase complex subunit E1p gene in Corynebacterium glutamicum" 16 : 236-239, 2009

      16 Süsstrunk U, "Pleiotropic effects of cAMP on germination, antibiotic biosynthesis and morphological development in Streptomyces coelicolor" 30 : 33-46, 1998

      17 Sambrook J, "Molecular Cloning:A Laboratory Manual" Cold Spring Harbor Laboratory Press 1989

      18 박수동, "Isolation and Characterization of TranscriptionalElements from Corynebacterium glutamicum" 한국미생물·생명공학회 14 (14): 789-795, 2004

      19 Toyoda K, "Involvement of the LuxR-type transcriptional regulator RamA in regulation of expression of the gapA gene, encoding glyceraldehyde-3-phosphate dehydrogenase of Corynebacterium glutamicum" 191 : 968-977, 2009

      20 Toyoda K, "Involvement of regulatory interactions among global regulators GlxR, SugR, and RamA in expression of ramA in Corynebacterium glutamicum" 195 : 1718-1726, 2013

      21 Liberman E, "Inhibition of E. coli adenylate cyclase activity by inorganic orthophosphate is dependent on IIIglc of the phosphoenolpyruvate:glycose phosphotransferase system" 141 : 1138-1144, 1986

      22 Sindelar G, "Improving lysine production by Corynebacterium glutamicum through DNA microarraybased identification of novel target genes" 76 : 677-689, 2007

      23 Cramer A, "Identification of RamA, a novel LuxR-type transcriptional regulator of genes involved in acetate metabolism of Corynebacterium glutamicum" 188 : 2554-2567, 2006

      24 Kim HJ, "Identification and characterization of glxR, a gene involved in regulation of glyoxylate bypass in Corynebacterium glutamicum" 186 : 3453-3460, 2004

      25 Cho HY, "Identification and characterization of a transcriptional regulator, SucR, that influences sucCD transcription in Corynebacterium glutamicum" 401 : 300-305, 2010

      26 Jungwirth B, "High-resolution detection of DNA binding sites of the global transcriptional regulator GlxR in Corynebacterium glutamicum" 159 : 12-22, 2013

      27 Schäfer A, "High-frequency conjugal plasmid transfer from gram-negative Escherichia coli to various gram-positive coryneform bacteria" 172 : 1663-1666, 1990

      28 Eggeling L, "Handbook of Corynebacterium glutamicum" CRC Press 535-566, 2005

      29 Toyoda K, "Genomewide identification of in vivo binding sites of GlxR, a cyclic AMP receptor protein-type regulator in Corynebacterium glutamicum" 193 : 4123-4133, 2011

      30 Park SY, "Functional characterization of the glxR deletion mutant of Corynebacterium glutamicum ATCC 13032: involvement of GlxR in acetate metabolism and carbon catabolite repression" 304 : 107-115, 2010

      31 Arndt A, "Ethanol catabolism in Corynebacterium glutamicum" 15 : 222-233, 2008

      32 Petersen S, "Essential role for cyclic AMP and its receptor protein in Yersinia enterocolitica virulence" 70 : 3665-3672, 2002

      33 Bindu Subhadra, "Elucidation of the Regulation of Ethanol Catabolic Genes and ptsG Using a glxR and Adenylate Cyclase Gene (cyaB) Deletion Mutants of Corynebacterium glutamicum ATCC 13032" 한국미생물·생명공학회 23 (23): 1683-1690, 2013

      34 Tauch A, "Efficient electrotransformation of Corynebacterium diphtheriae with a mini-replicon derived from the Corynebacterium glutamicum plasmid pGA1" 45 : 362-367, 2002

      35 Han SO, "Effect of carbon source availability and growth phase on expression of Corynebacterium glutamicum genes involved in the tricarboxylic acid cycle and glyoxylate bypass" 154 : 3073-3083, 2008

      36 Schreiner ME, "E1 enzyme of the pyruvate dehydrogenase complex in Corynebacterium glutamicum: molecular analysis of the gene and phylogenetic aspects" 187 : 6005-6018, 2005

      37 Auchter M, "Dual transcriptional control of the acetaldehyde dehydrogenase gene ald of Corynebacterium glutamicum by RamA and RamB" 140 : 84-91, 2009

      38 Botsford JL, "Cyclic AMP in prokaryotes" 56 : 100-122, 1992

      39 Arndt A, "Corynebacteria: Genomics and Molecular Biology" Caister Academic Press 155-182, 2008

      40 Baumbach J, "CoryneRegNet 4.0 - A reference database for corynebacterial gene regulatory networks" 8 : 429-, 2007

      41 Auchter M, "Control of adhA and sucR expression by the SucR regulator in Corynebacterium glutamicum" 152 : 77-86, 2011

      42 Yukawa H, "Comparative analysis of the Corynebacterium glutamicum group and complete genome sequence of strain R" 153 : 1042-1058, 2007

      43 Frunzke J, "Co-ordinated regulation of gluconate catabolism and glucose uptake in Corynebacterium glutamicum by two functionally equivalent transcriptional regulators, GntR1 and GntR2" 67 : 305-322, 2008

      44 Ooyen J, "Citrate synthase in Corynebacterium glutamicum is encoded by two gltA transcripts which are controlled by RamA, RamB, and GlxR" 154 : 140-148, 2011

      45 Shaw WV, "Chloramphenicol acetyltransferase from chloramphenicol-resistant bacteria" 43 : 737-755, 1975

      46 Cha PH, "Characterization of an adenylate cyclase gene (cyaB) deletion mutant of Corynebacterium glutamicum ATCC 13032" 85 : 1061-1068, 2010

      47 Letek M, "Characterization and use of catabolite-repressed promoters from gluconate genes in Corynebacterium glutamicum" 188 : 409-423, 2006

      48 Brückner R, "Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization" 209 : 141-148, 2002

      49 Ikeda M., "Amino acid production processes" 79 : 1-35, 2003

      50 Simon R, "A broad host range vector mobilization system for in vivo genetic engineering:transposon mutagenesis in gram-negative bacteria" 1 : 784-791, 1983

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