This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Handke, L. D.
Right arrow Articles by Sonenshein, A. L.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Handke, L. D.
Right arrow Articles by Sonenshein, A. L.

 Previous Article  |  Next Article 

Journal of Bacteriology, February 2008, p. 798-806, Vol. 190, No. 3
0021-9193/08/$08.00+0     doi:10.1128/JB.01115-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Interaction of Bacillus subtilis CodY with GTP{triangledown} ,{dagger}

Luke D. Handke,1 Robert P. Shivers,1,2,{ddagger} and Abraham L. Sonenshein1,2*

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

Received 16 July 2007/ Accepted 5 November 2007

Many of the adaptive mechanisms that allow Bacillus subtilis to adjust to changes in nutrient availability are controlled by CodY. Binding of CodY to its target genes is stimulated by interaction with its effectors, GTP and the branched-chain amino acids (BCAAs). Upon nutrient limitation, intracellular pools of these effectors are depleted and CodY can no longer repress genes required for adaptation. In vitro studies reported here explored in more detail the interaction of CodY with GTP. DNase I footprinting experiments indicated that CodY has an affinity for GTP in the millimolar range. Further, CodY was shown to interact specifically with GTP and dGTP; no other naturally occurring nucleotides that were tested, including ppGpp and pppGpp, resulted in DNA protection. Two nonhydrolyzable analogs of GTP were fully able to activate CodY binding to target DNA, demonstrating that GTP hydrolysis is not necessary for CodY-dependent regulation. GTP and the BCAAs were shown to act additively to increase the affinity of CodY for DNA; increased protection was observed in DNase I footprinting experiments when both effectors were present, compared to either effector alone, and in in vitro transcription reactions, transcriptional repression by CodY was stronger in the presence of both GTP and BCAAs than of BCAAs alone. Thus, interaction of CodY with GTP is specific and results in increased affinity for its target genes. This increase in affinity is independent of GTP hydrolysis and is augmented in the presence of BCAAs.

* 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 9 November 2007.

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

{ddagger} Present address: Department of Biology, Massachusetts Institute of Technology, Cambridge, MA 02139.

Journal of Bacteriology, February 2008, p. 798-806, Vol. 190, No. 3
0021-9193/08/$08.00+0     doi:10.1128/JB.01115-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Somerville, G. A., Proctor, R. A. (2009). At the Crossroads of Bacterial Metabolism and Virulence Factor Synthesis in Staphylococci. Microbiol. Mol. Biol. Rev. 73: 233-248 [Abstract] [Full Text]  
  • Tzeng, Y.-L. (2009). A Unique Sensor Histidine Kinase. J. Bacteriol. 191: 681-682 [Full Text]  
  • Lemos, J. A., Nascimento, M. M., Lin, V. K., Abranches, J., Burne, R. A. (2008). Global Regulation by (p)ppGpp and CodY in Streptococcus mutans. J. Bacteriol. 190: 5291-5299 [Abstract] [Full Text]  
  • Belitsky, B. R., Sonenshein, A. L. (2008). Genetic and Biochemical Analysis of CodY-Binding Sites in Bacillus subtilis. J. Bacteriol. 190: 1224-1236 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»