Previous Article | Next Article ![]()
Journal of Bacteriology, February 2006, p. 1411-1418, Vol. 188, No. 4
0021-9193/06/$08.00+0 doi:10.1128/JB.188.4.1411-1418.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.
Amrita Kumar,
Travis H. Wyman,
and
Charles P. Moran Jr.*
Department of Microbiology and Immunology, Emory University School of Medicine, Atlanta, Georgia 30322
Received 19 August 2005/ Accepted 30 November 2005
At the onset of endospore formation in Bacillus subtilis the DNA-binding protein Spo0A directly activates transcription from promoters of about 40 genes. One of these promoters, Pskf, controls expression of an operon encoding a killing factor that acts on sibling cells. AbrB-mediated repression of Pskf provides one level of security ensuring that this promoter is not activated prematurely. However, Spo0A also appears to activate the promoter directly, since Spo0A is required for Pskf activity in a
abrB strain. Here we investigate the mechanism of Pskf activation. DNase I footprinting was used to determine the locations at which Spo0A bound to the promoter, and mutations in these sites were found to significantly reduce promoter activity. The sequence near the 10 region of the promoter was found to be similar to those of extended 10 region promoters, which contain a TRTGn motif. Mutational analysis showed that this extended 10 region, as well as other base pairs in the 10 region, is required for Spo0A-dependent activation of the promoter. We found that a substitution of the consensus base pair for the nonconsensus base pair at position 9 of Pskf produced a promoter that was active constitutively in both
abrB and
spo0A
abrB strains. Therefore, the base pair at position 9 of Pskf makes its activity dependent on Spo0A binding, and the extended 10 region motif of the promoter contributes to its high level of activity.
G.C., A.K., and T.H.W. contributed equally to this work.
This article has been cited by other articles:
Copyright © 2009 by the American Society for Microbiology. For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»