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Originally published In Press as doi:10.1074/jbc.M009864200 on December 15, 2000
J. Biol. Chem., Vol. 276, Issue 13, 9620-9625, March 30, 2001
Roles of Glucitol in the GutR-mediated Transcription Activation
Process in Bacillus subtilis
TIGHT BINDING OF GutR TO ITS BINDING SITE*
Karen K. H.
Poon ,
Chyi-Liang
Chen, and
Sui-Lam
Wong§
From the Division of Cellular, Molecular and Microbial Biology,
Department of Biological Sciences, University of Calgary, Calgary,
Alberta T2N 1N4, Canada
Glucitol induction in Bacillus
subtilis requires a transcription activator, GutR, and a sequence
located upstream of the gut promoter. To understand the
initial steps involved in the GutR-mediated transcription activation
process and the physiological roles of glucitol, GutR was overproduced
and purified. In the absence of glucitol, GutR exists as a monomer and
binds directly to its binding site in the gut regulatory
region. This binding site was mapped to a 29-base pair imperfect
inverted repeat located between 78 and 50, and there is only one
GutR binding site within the regulatory region. The kinetic parameters
of the interaction between GutR and its binding site were monitored in
real time using surface plasmon resonance. The half-life of the
GutR-DNA complex in the absence of glucitol was estimated to be 6.8 min. In contrast, in the presence of glucitol, the half-life of the
complex was extended to longer than 19 h by affecting only the
off-rate but not the on-rate. This effect is glucitol-specific. These
data indicate that glucitol binds to GutR and induces GutR to have an
extremely tight binding at its binding site. The physiological relevance of this process in transcription activation is discussed.
*
This project is supported by a research grant from the
Natural Science and Engineering Research Council of Canada (to
S. L. W.).The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Supported in part by a research assistantship from the Department
of Biological Sciences at the University of Calgary. Current address:
University of Guelph, Dept. of Microbiology, Guelph, Ontario N1G 2W1, Canada.
§
To whom correspondence should be addressed. Tel.:
403-220-5721; Fax: 403-289-9311; E-mail: slwong@ucalgary.ca.
Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.

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Copyright © 2001 by the American Society for Biochemistry and Molecular Biology.
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