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(Received for publication, May 1, 1996, and in revised form, August 22, 1996)
From the Saccharomyces cerevisiae contains two
structural genes, ACS1 and ACS2, each encoding
an active acetyl-coenzyme A synthetase. Characterization of enzyme
activities in cell-free extracts from strains expressing either of the
two genes revealed differences in the catalytic properties of the two
enzymes. The Km for acetate of Acs1p was about
30-fold lower than that of Acs2p and Acs1p, but not Acs2p, could use
propionate as a substrate. Enzyme activity measurements and mRNA
analyses showed that ACS1 and ACS2 were both
expressed during carbon-limited growth on glucose, ethanol, and acetate
in aerobic chemostat cultures. In anaerobic glucose-limited cultures,
only the ACS2 gene was expressed. Based on these facts, the
products of the ACS1 and ACS2 genes were
identified as the previously described "aerobic" and
"non-aerobic" forms of acetyl-coenzyme A synthetase, respectively.
Batch and glucose-pulse experiments revealed that transcription of
ACS1 is subject to glucose repression. A mutant strain
lacking Acs2p was unable to grow on glucose in batch cultures, but grew
readily in aerobic glucose-limited chemostat cultures, in which the low
residual glucose concentration alleviated glucose repression.
Experiments in which ethanol was pulsed to aerobic ethanol-limited
chemostat cultures indicated that, in addition to glucose, ethanol also repressed ACS1 transcription, although to a lesser extent.
In contrast, transcription of ACS2 was slightly induced by
ethanol and glucose. Absence of ACS2 prevented complete
glucose repression of ACS1, indicating that
ACS2 (in)directly is involved in the transcriptional
regulation of ACS1.
Volume 271, Number 46,
Issue of November 15, 1996
pp. 28953-28959
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
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Kluyver Laboratory of Biotechnology,
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