Nucleic Acids Research, Vol 27, Issue 22 4391-4398, Copyright © 1999 by Oxford University Press
M Grauslund, JM Lopes and B Ronnow
In Saccharomyces cerevisiae glycerol utilization is mediated by two
enzymes, glycerol kinase (Gut1p) and mitochondrial glycerol-3-phosphate
dehydrogenase (Gut2p). The carbon source regulation of GUT1 was studied
using promoter-reporter gene fusions. The promoter activity was lowest
during growth on glucose and highest on the non-fermentable carbon sources,
glycerol, ethanol, lactate, acetate and oleic acid. Mutational analysis of
the GUT1 promoter region showed that two upstream activation sequences,
UAS(INO) and UAS(ADR1), are responsible for approximately 90% of the
expression during growth on glycerol. UAS(ADR1) is a presumed binding site
for the zinc finger transcription factor Adr1p and UAS(INO) is a presumed
binding site for the basic helix-loop-helix transcription factors Ino2p and
Ino4p. In vitro experiments showed Adr1 and Ino2/Ino4 protein-dependent
binding to UAS(ADR1) and UAS(INO). The negative regulator Opi1p mediates
repression of the GUT1 promoter, whereas the effects of the glucose
repressors Mig1p and Mig2p are minor. Together, the experiments show that
GUT1 is carbon source regulated by different activation and repression
systems.
ARTICLES
Expression of GUT1, which encodes glycerol kinase in Saccharomyces cerevisiae, is controlled by the positive regulators Adr1p, Ino2p and Ino4p and the negative regulator Opi1p in a carbon source-dependent fashion
Danisco Biotechnology, Danisco A/S, Langebrogade 1, DK-1001 Copenhagen K, Denmark.
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