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J Biol Chem, Vol. 275, Issue 19, 14031-14037, May 12, 2000
From the Bacillus subtilis possesses two
similar putative phosphorylating glyceraldehyde-3-phosphate
dehydrogenase (GAPDH) encoding genes, gap (renamed
gapA) and gapB. A gapA mutant was
unable to grow on glycolytic carbon sources, although it developed as
well as the wild-type strain on gluconeogenic carbon sources. A
gapB mutant showed the opposite phenotype. Purified GapB
showed a 50-fold higher GAPDHase activity with NADP+ than
with NAD+, with Km values of 0.86 and
5.7 mM, respectively. lacZ reporter gene
fusions revealed that the gapB gene is transcribed during
gluconeogenesis and repressed during glycolysis. Conversely, gapA transcription is 5-fold higher under glycolytic
conditions than during gluconeogenesis. GAPDH activity assays in crude
extracts of wild-type and mutant strains confirmed this differential
expression pattern at the enzymatic level. Genetic analyses
demonstrated that gapA transcription is repressed by the
yvbQ (renamed cggR) gene product and indirectly
stimulated by CcpA. Thus, the same enzymatic step is catalyzed in
B. subtilis by two enzymes specialized, through the
regulation of their synthesis and their enzymatic characteristics,
either in catabolism (GapA) or in anabolism (GapB). Such a dual
enzymatic system for this step of the central carbon metabolism is
described for the first time in a nonphotosynthetic eubacterium, but
genomic analyses suggest that it could be a widespread feature.
Two Glyceraldehyde-3-phosphate Dehydrogenases with Opposite
Physiological Roles in a Nonphotosynthetic Bacterium*
§,
,**
Génétique Moléculaire et
Cellulaire, INRA-CNRS (URA1925), 78850 Thiverval-Grignon, France,
¶ UMR7567-CNRS-UHP-Maturation des ARN et Enzymologie
Moléculaire, Faculté des Sciences, Bld des Aiguillettes,
BP239, 54506 Vandoeuvre-les-Nançy, France, and the
Génétique Microbienne, INRA,
78352 Jouy-en-Josas, France
*
This work was supported by the EU Biotechnology Program
Grant BIO-4CT95-0278.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 © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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