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J. Biol. Chem., Vol. 278, Issue 4, 2169-2176, January 24, 2003
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From the We found that a polycistronic operon
(ywfBCDEFG) and a monocistronic gene (ywfH) are
required for the biosynthesis of bacilysin in Bacillus
subtilis. The disruption of these genes by plasmid integration
caused loss of the ability to produce bacilysin, accompanied by a lack
of bacilysin synthetase activity in the crude extract. We investigated
the regulatory mechanism for bacilysin biosynthesis using the
transcriptional lacZ fusion system. The transcription of
these genes was found to be induced at the transition from exponential
to stationary phase. Induction of transcription was accelerated by
depleting a required amino acid, which was done by transferring the
wild-type (rel+) cells to an amino acid-limited
medium. In contrast, no enhancement of the gene expression was detected
in relA mutant cells. In wild-type (rel+) cells, a forced reduction of
intracellular GTP, brought about by addition of decoyinine, which is a
GMP synthetase inhibitor, enhanced the expression of both the
ywfBCDEFG operon and the ywfH gene, resulting
in a 2.5-fold increase in bacilysin production. Disruption of the
codY gene, which regulates stationary phase genes by
detecting the level of GTP, also induced transcription of these genes.
In contrast, the expression of ywfBCDEFG in
relA cells was not activated either by decoyinine addition
or codY disruption, although the expression of
ywfH was induced. Moreover, the codY disruption
resulted in an increase of bacilysin production only in
rel+ cells. These results indicate that
guanosine 5'-diphosphate 3'-diphosphate (ppGpp) plays a crucial role in
transcription of the ywfBCDEFG operon and that the
transcription of these genes are dependent upon the level of
intracellular GTP which is transmitted as a signal via the
CodY-mediated repression system. We propose that, unlike antibiotic
production in Streptomyces spp., bacilysin production in
B. subtilis is controlled by a dual regulation
system composed of the guanine nucleotides ppGpp and GTP.
Guanine Nucleotides Guanosine 5'-Diphosphate 3'-Diphosphate
and GTP Co-operatively Regulate the Production of an Antibiotic
Bacilysin in Bacillus subtilis*
,,¶
Microbial Function Laboratory and
§ Molecular Elucidation Laboratory, National Food Research
Institute, Tsukuba, Ibaraki 305-8642, Japan
*
This work was supported by a grant from the Organized
Research Combination System of the Science and Technology Agency of Japan.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.
Copyright © 2003 by The American Society for Biochemistry and Molecular Biology, Inc.
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