Cécile Jourlin-Castelli

Publications (3)13.01 Total impact

• Article: Mechanism of repression by Bacillus subtilis CcpC, a LysR family regulator.

  Sam-In Kim, Cécile Jourlin-Castelli, Stephen R Wellington, Abraham L Sonenshein
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  ABSTRACT: Bacillus subtilis CcpC is a LysR family transcriptional regulatory protein that negatively regulates genes encoding enzymes of the tricarboxylic acid branch of the Krebs cycle. In the present work, the promoter region of the aconitase (citB) gene was used to investigate the mechanism of repression by CcpC. The binding of CcpC to the citB promoter region was shown to depend on DNA elements located near positions -66 and -27. Binding to these elements induced a bend in the DNA at position -41. Introduction of mutations in the -27 region and the presence of citrate, the inducer, had similar effects. In either case, citB expression was derepressed in vivo, the affinity of CcpC binding was reduced in vitro, the angle of the bend was relaxed, and RNA polymerase gained greater access to the -35 region of the promoter.
  Journal of Molecular Biology 01/2004; 334(4):609-24. · 4.00 Impact Factor
• Article: Regulation of the bacillus subtilis ccpC gene by ccpA and ccpC.

  Hyun-Jin Kim, Cécile Jourlin-Castelli, Sam-In Kim, Abraham L Sonenshein
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  ABSTRACT: Bacillus subtilis CcpC, a LysR-type transcriptional regulator, represses the transcription of genes for citrate synthase (citZ) and aconitase (citB) in response to citrate availability. Transcription of ccpC was shown to initiate at two promoters, P1, located just upstream of the ccpC gene, and P2, located within or upstream of the neighbouring ykuL gene. Expression from the ccpC-specific promoter (P1) was negatively regulated by CcpC but independent of the carbon source in the medium. Gel shift and DNase I footprinting experiments revealed that CcpC binds to an interrupted dyad sequence that surrounds the ccpC transcriptional start point. Transcription of ccpC from the upstream promoter (P2) was repressed by glucose in a CcpA-dependent manner. A putative CcpA binding site (cre) was identified upstream of the -35 region of the P1 promoter. Transcriptional fusion studies demonstrated that glucose repression of ccpC expression from the P2 promoter depends on this cre site. In addition, DNase I footprinting experiments showed that CcpA specifically binds to this cre site and that the introduction of mutations (cre*) into this site abolished the binding. These results suggest that CcpA may control CcpC synthesis by acting as a road-block to readthrough transcription from the P2 promoter.
  Molecular Microbiology 02/2002; 43(2):399-410. · 5.01 Impact Factor
• Article: CcpC, a novel regulator of the LysR family required for glucose repression of the citB gene in Bacillus subtilis

  Cécile Jourlin-Castelli, Nagraj Mani, Michiko M Nakano, Abraham L Sonenshein
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  ABSTRACT: Synergistic carbon catabolite repression of the Bacillus subtilis aconitase (citB) gene by glucose and a source of 2-ketoglutarate is dependent on DNA sequences located upstream of the gene. Mutations in a dyad symmetry element centered at position −66 and in a repeat of the downstream arm of the dyad symmetry at position −27 cause derepressed citB expression. In this work, a protein able to bind to a DNA fragment containing these elements was purified and identified. This protein, named CcpC (Catabolite control protein C), shares sequence similarity with members of the LysR family of transcriptional regulators. In addition to binding to the citB promoter, CcpC bound to the promoter of the citZ gene, which encodes the cell’s major citrate synthase and is subject to carbon catabolite repression. In a ccpC null mutant, expression of both citB and citZ was derepressed in glucose-glutamine minimal medium, indicating that CcpC is a negative regulator of citB and citZ gene expression. DNase I footprinting experiments showed that CcpC binds to two sites within the citB promoter region, corresponding to the dyad symmetry and −27 elements. In the presence of citrate, a putative inducer, only the dyad symmetry element was fully protected by CcpC. When the dyad symmetry element was mutated, CcpC was no longer able to bind to either the dyad symmetry or −27 elements. Repression of citB and citZ gene expression during anaerobiosis also proved to be mediated by CcpC.
  Journal of Molecular Biology 02/2000; · 4.00 Impact Factor