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(Received for publication, February 29, 1996, and in revised form, July 23, 1996)
From the The Saccharomyces cerevisiae PIS1
gene encodes phosphatidylinositol synthase. The amount of
phosphatidylinositol synthase is not affected by the presence of
inositol and choline in the growth medium. This is unusual because the
amounts and/or activities of other phospholipid biosynthetic enzymes
are affected by these precursors, and the promoter of the PIS1
gene contains a sequence resembling the regulatory element that
coordinates the inositol-mediated regulation (UASINO).
We found that transcription of the PIS1 gene was
insensitive to inositol and choline and did not require the putative
UASINO regulatory sequence or the cognate regulatory genes
(INO2 and OPI1).
The PIS1 promoter includes sequences (MCEs) that bind the
Mcm1 protein. Because the Mcm1 protein interacts with both the Sln1 and
the Gal11 regulatory proteins, we examined the effect of mutant alleles
of the MCM1 and SLN1 genes and carbon source on
expression of the PIS1 gene. We found that expression of
the PIS1 gene was reduced when cells were grown in a medium
containing glycerol and increased when grown in a medium containing
galactose relative to cells grown in a glucose medium. The
glycerol-mediated repression of PIS1 gene expression
required both the MCM1 gene and the MCEs, whereas the
SLN1 gene was required for full galactose-mediated
induction of a PIS1-lacZ reporter gene.
Thus, PIS1 gene expression is unique among the phospholipid
biosynthetic structural genes because it is uncoupled from the inositol
response and regulated in response to the carbon source. This is the
first example in yeast of a complete circuit linking a stimulus (carbon
source) to gene regulation (PIS1) using a two-component
regulator (SLN1).
Volume 271, Number 43,
Issue of October 25, 1996
pp. 26596-26601
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
and
¶
Department of Molecular and Cellular
Biochemistry and the ¶ Program in Molecular Biology, Loyola
University of Chicago, Maywood, Illinois 60153
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