NDLI: 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 ▿

Content Provider	PubMed Central
Author	Susanna, Kim A. Mironczuk, Aleksandra M. Smits, Wiep Klaas Hamoen, Leendert W. Kuipers, Oscar P.
Copyright Year	2007
Abstract	The competence transcription factor ComK plays a central role in competence development in Bacillus subtilis by activating the transcription of the K regulon. ComK-activated genes are characterized by the presence of a specific sequence to which ComK binds, a K-box, in their upstream DNA region. Each K-box consists of two AT-boxes with the consensus sequence AAAA-(N)5-TTTT, which are separated by a flexible spacer resulting in either two, three, or four helical turns between the starting nucleotides of the repeating AT-box units. In this study, the effects of potential determinants of ComK regulation in K-boxes were investigated by testing ComK's transcription activation and DNA-binding affinity on altered K-boxes with mutations either in the spacer between the AT-boxes or in the consensus sequence of the AT-boxes. The most striking result demonstrates the importance of the second thymine base in the AT-boxes. Mutation of this T into a guanine resulted in a threefold reduction in transcription activation and DNA binding by ComK. Transcription activation, as well as DNA binding, was almost completely abolished when both AT-boxes contained a T2-to-G mutation. This result was corroborated by in silico analyses demonstrating that a combination of mutations at the T2 positions of both AT-boxes is not found among any ComK-activated K-boxes, indicating that at least one consensus T2 position is required to maintain a functional K-box. The results suggest an important structural role for T2 in ComK binding, probably by its specific position in the minor groove of the DNA.
Related Links	http://dx.doi.org/10.1128/jb.00281-07
Ending Page	4728
Page Count	11
Starting Page	4718
File Format	PDF
ISSN	00219193
e-ISSN	10985530
Journal	Journal of Bacteriology
Issue Number	13
Volume Number	189
Language	English
Publisher	American Society for Microbiology
Publisher Date	2007-07-01
Access Restriction	Open
Rights Holder	American Society for Microbiology
Subject Keyword	Molecular Biology Microbiology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Microbiology

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