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Journal of Bacteriology, February 2004, p. 1120-1128, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1120-1128.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Mechanism of Transcription Activation at the comG Promoter by the Competence Transcription Factor ComK of Bacillus subtilis

K. A. Susanna,1 A. F. van der Werff,1,{dagger} C. D. den Hengst,1 B. Calles,2 M. Salas,2 G. Venema,1 L. W. Hamoen,1* and O. P. Kuipers1*

Department of Genetics, University of Groningen, NL-9751 NN Haren, The Netherlands,1 Centro de Biologia Molecular "Servero Ochoa" (CSIC-UAM), Universidad Autonoma, Cantoblanco 28049, Madrid, Spain2

Received 2 July 2003/ Accepted 13 October 2003

The development of genetic competence in Bacillus subtilis is regulated by a complex signal transduction cascade, which results in the synthesis of the competence transcription factor, encoded by comK. ComK is required for the transcription of the late competence genes that encode the DNA binding and uptake machinery and of genes required for homologous recombination. In vivo and in vitro experiments have shown that ComK is responsible for transcription activation at the comG promoter. In this study, we investigated the mechanism of this transcription activation. The intrinsic binding characteristics of RNA polymerase with and without ComK at the comG promoter were determined, demonstrating that ComK stabilizes the binding of RNA polymerase to the comG promoter. This stabilization probably occurs through interactions with the upstream DNA, since a deletion of the upstream DNA resulted in an almost complete abolishment of stabilization of RNA polymerase binding. Furthermore, a strong requirement for the presence of an extra AT box in addition to the common ComK-binding site was shown. In vitro transcription with B. subtilis RNA polymerase reconstituted with wild-type {alpha}-subunits and with C-terminal deletion mutants of the {alpha}-subunits was performed, demonstrating that these deletions do not abolish transcription activation by ComK. This indicates that ComK is not a type I activator. We also show that ComK is not required for open complex formation. A possible mechanism for transcription activation is proposed, implying that the major stimulatory effect of ComK is on binding of RNA polymerase.

* Corresponding author. Mailing address for L. W. Hamoen: Sir William Dunn School of Pathology, University of Oxford, South Parks Rd., Oxford OX1 3RE, United Kingdom. E-mail: L.W.Hamoen{at}biol.rug.nl. Mailing address for O. P. Kuipers: Department of Genetics, University of Groningen, NL-9751 NN Haren, The Netherlands. Phone: 31(0)50.363.2093. Fax: 31(0)50.363.2348. E-mail: O.P.Kuipers{at}biol.rug.nl.

{dagger} Present address: Unilever Bestfoods Nederland B.V., NL-3071 JL Rotterdam, The Netherlands.

Journal of Bacteriology, February 2004, p. 1120-1128, Vol. 186, No. 4
0021-9193/04/$08.00+0     DOI: 10.1128/JB.186.4.1120-1128.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Susanna, K. A., Mironczuk, A. M., Smits, W. K., Hamoen, L. W., Kuipers, O. P. (2007). A Single, Specific Thymine Mutation in the ComK-Binding Site Severely Decreases Binding and Transcription Activation by the Competence Transcription Factor ComK of Bacillus subtilis. J. Bacteriol. 189: 4718-4728 [Abstract] [Full Text]  
  • Susanna, K. A., Fusetti, F., Thunnissen, A.-M. W. H., Hamoen, L. W., Kuipers, O. P. (2006). Functional analysis of the competence transcription factor ComK of Bacillus subtilis by characterization of truncation variants. Microbiology 152: 473-483 [Abstract] [Full Text]  
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