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Journal of Bacteriology, December 1998, p. 6681-6688, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
The First Gene of the Bacillus subtilis clpC Operon,
ctsR, Encodes a Negative Regulator of Its Own
Operon and Other Class III Heat Shock Genes
Elke
Krüger and
Michael
Hecker*
Institut für Mikrobiologie und
Molekularbiologie, Ernst-Moritz-Arndt-Universität, D-17487
Greifswald, Germany
Received 3 August 1998/Accepted 6 October 1998
The Bacillus subtilis clpC operon is regulated by two
stress induction pathways relying on either
B or a class
III stress induction mechanism acting at a
A-like
promoter. When the clpC operon was placed under the
control of the isopropyl-
-D-thiogalactopyranoside
(IPTG)-inducible Pspac promoter, dramatic
repression of the natural clpC promoters fused to a
lacZ reporter gene was noticed after IPTG induction. This result strongly indicated negative regulation of the clpC
operon by one of its gene products. Indeed, the negative regulator
could be identified which is encoded by the first gene of the
clpC operon, ctsR, containing a predicted
helix-turn-helix DNA-binding motif. Deletion of ctsR
abolished the negative regulation and resulted in high expression of
both the clpC operon and the clpP gene under nonstressed conditions. Nevertheless, a further increase in
clpC and clpP mRNA levels was observed after
heat shock, even in the absence of
B, suggesting a
second induction mechanism at the vegetative promoter. Two-dimensional
gel analysis and mRNA studies showed that the expression of other class
III stress genes was at least partially influenced by the
ctsR deletion. Studies with different clpC
promoter fragments either fused to the reporter gene bgaB
or used in gel mobility shift experiments with the purified CtsR
protein revealed a possible target region where the repressor seemed to
bind in vivo and in vitro. Our data demonstrate that the CtsR protein acts as a global repressor of the clpC operon, as well as
other class III heat shock genes, by preventing unstressed
transcription from either the
B- or
A-dependent promoter and might be inactivated or
dissociate under inducing stress conditions.
*
Corresponding author. Mailing address: Institut
für
Mikrobiologie und Molekularbiologie, Ernst-Moritz-Arndt-Universität, D-17487 Greifswald,
Germany. Phone: 03834/864200. Fax: 03834/864202. E-mail: hecker{at}microbio7.biologie.uni-greifswald.de.
Journal of Bacteriology, December 1998, p. 6681-6688, Vol. 180, No. 24
0021-9193/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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