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Journal of Bacteriology, February 2009, p. 862-872, Vol. 191, No. 3
0021-9193/09/$08.00+0     doi:10.1128/JB.01384-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

CcpC-Dependent Regulation of Citrate Synthase Gene Expression in Listeria monocytogenes{triangledown}

Meghna Mittal,1 Silvia Picossi,2,{dagger} and Abraham L. Sonenshein1,2*

Program in Molecular Microbiology, Sackler School of Graduate Biomedical Sciences,1 Department of Molecular Biology and Microbiology, School of Medicine, Tufts University, Boston, Massachusetts 021112

Received 2 October 2008/ Accepted 4 November 2008

Citrate synthase, the first and rate-limiting enzyme of the tricarboxylic acid branch of the Krebs cycle, was shown to be required for de novo synthesis of glutamate and glutamine in Listeria monocytogenes. The citrate synthase (citZ) gene was found to be part of a complex operon with the upstream genes lmo1569 and lmo1568. The downstream isocitrate dehydrogenase (citC) gene appears to be part of the same operon as well. Two promoters were shown to drive citZ expression, a distal promoter located upstream of lmo1569 and a proximal promoter located upstream of the lmo1568 gene. Transcription of citZ from both promoters was regulated by CcpC by interaction with a single site; assays of transcription in vivo and assays of CcpC binding in vitro revealed that CcpC interacts with and represses the proximal promoter that drives expression of the lmo1568, citZ, and citC genes and, by binding to the same site, prevents read-through transcription from the distal, lmo1569 promoter. Expression of the lmo1568 operon was not affected by the carbon source but was repressed during growth in complex medium by addition of glutamine.

* Corresponding author. Mailing address: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Avenue, Boston, MA 02111. Phone: (617) 636-6761. Fax: (617) 636-0337. E-mail: linc.sonenshein{at}tufts.edu

{triangledown} Published ahead of print on 14 November 2008.

{dagger} Present address: Instituto de Bioquímica Vegetal y Fotosíntesis, Consejo Superior de Investigaciones Científicas and Universidad de Sevilla, E-41092 Seville, Spain.

Journal of Bacteriology, February 2009, p. 862-872, Vol. 191, No. 3
0021-9193/09/$08.00+0     doi:10.1128/JB.01384-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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