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Journal of Bacteriology, February 2008, p. 1224-1236, Vol. 190, No. 4
0021-9193/08/$08.00+0     doi:10.1128/JB.01780-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genetic and Biochemical Analysis of CodY-Binding Sites in Bacillus subtilis{triangledown} ,{dagger}

Boris R. Belitsky* and Abraham L. Sonenshein

Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111

Received 9 November 2007/ Accepted 6 December 2007

CodY is a global transcriptional regulator that is known to control directly the expression of at least two dozen operons in Bacillus subtilis, but the rules that govern the binding of CodY to its target DNA have been unclear. Using DNase I footprinting experiments, we identified CodY-binding sites upstream of the B. subtilis ylmA and yurP genes. The protected regions overlapped versions of a previously proposed CodY-binding consensus motif, AATTTTCWGAAAATT. Multiple single mutations were introduced into the CodY-binding sites of the ylmA, yurP, dppA, and ilvB genes. The mutations affected both the affinity of CodY for its binding sites in vitro and the expression in vivo of lacZ fusions that carry these mutations in their promoter regions. Our results show that versions of the AATTTTCWGAAAATT motif, first identified for Lactococcus lactis CodY, with up to five mismatches play an important role in the interaction of B. subtilis CodY with DNA.

* Corresponding author. Mailing address: Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Ave., Boston, MA 02111. Phone: (617) 636-3618. Fax: (617) 636-0337. E-mail: Boris.Belitsky{at}tufts.edu

{triangledown} Published ahead of print on 14 December 2007.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, February 2008, p. 1224-1236, Vol. 190, No. 4
0021-9193/08/$08.00+0     doi:10.1128/JB.01780-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Pohl, K., Francois, P., Stenz, L., Schlink, F., Geiger, T., Herbert, S., Goerke, C., Schrenzel, J., Wolz, C. (2009). CodY in Staphylococcus aureus: a Regulatory Link between Metabolism and Virulence Gene Expression. J. Bacteriol. 191: 2953-2963 [Abstract] [Full Text]  


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