Nucleic Acids Research, 1992, Vol. 20, No. 22 5955-5961
© 1992
MOLECULAR BIOLOGY |
Regulatory gene INO4 of yeast phospholipid biosynthesis is positively autoregulated and functions as a trans-activator of fatty acid synthase genes FAS1 and FAS2 from Saccharomyces cerevisiae
Institut f
r Mikrobiologie und Biochemie, Lehrstuhl Biochemie StaudtstraGe 5, D-8520 Erlangen, Germany
Received July 28, 1992. Revised October 23, 1992. Accepted October 23, 1992.
The sequence motif 5' TYTTCACATGY 3' functions as an upstream activation site common to both yeast fatty acid synthase genes, FAS1 and FAS2. In addition, this UASFAS element is shared by all so far characterized genes of yeast phospholipid biosynthesis. We have investigated the influence of a functional INO4 gene previously described as a regulator of inositol biosynthesis on the expression of FAS1 and FAS2. In a 
INO4 null allele strain, both genes are expressed at only 50% of wild type level. Using individual UASFAS sequence motifs inserted into a heterologous test system, a drastic decrease of reporter gene expression to 2 -10% of the wild type reference was observed in the AINO4 mutant. In gel retardation assays, the protein-DNA complex involving the previously described FAS binding factor 1, Fbf 1, was absent when using a protein extract from INO4 mutant. On the other hand, this signal was enhanced with an extract from cells grown under conditions of inositol/choline derepression. Subsequent experiments demonstrated that INO4 expression is itself affected by phospholipid precursors, mediated by an UASFAS element in the INO4 upstream region. Thus, in addition of being an activator of phospholipid biosynthetic genes, INO4 is also subject to a positive autoregulatory loop in its own biosynthesis.
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