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J. Biol. Chem., Vol. 276, Issue 46, 42901-42907, November 16, 2001

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The Bacillus subtilis Competence Transcription Factor, ComK, Overrides LexA-imposed Transcriptional Inhibition without Physically Displacing LexA^*

Leendert W. Hamoen^§, Bertjan Haijema^¶, Jetta J. Bijlsma, Gerard Venema, and Charles M. Lovett^**

From the  Department of Genetics, University of Groningen, NL-9751 NN Haren, The Netherlands, the ^¶ Institute of Virology, University of Utrecht, NL-3584 CL Utrecht, The Netherlands, the  Department of Medical Microbiology, Vrije Universiteit, Amsterdam, The Netherlands, and the ^** Department of Chemistry, Williams College, Williamstown, Massachusetts 01267

During the development of competence in Bacillus subtilis the recA gene is activated by the competence transcription factor, ComK, which is presumably required to alleviate the transcriptional repression of recA by LexA. To investigate the mechanism by which ComK activates recA transcription we examined the binding of ComK and LexA to the recA promoter in vitro. Using hydroxyl radical protection analyses to establish the location of ComK dimer-binding sites within the recA promoter, we identified four AT-boxes in a configuration unique for ComK-regulated promoters. Gel mobility shift experiments showed that all four ComK dimer-binding sites were occupied at ComK concentrations in the physiological range. In addition, occupation of all ComK-binding sites did not prevent LexA from binding to the recA promoter, despite the fact that the ComK and LexA recognition motifs partially overlap. Although ComK did not replace LexA from the recA promoter, in vitro transcription analyses indicated that the presence of ComK is sufficient to alleviate LexA repression of recA.

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