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J. Biol. Chem., Vol. 264, Issue 28, 16537-16544, Oct, 1989
JE Stukey, VM McDonough and CE Martin
The unsaturated fatty acid (ufa) requiring ole1 mutant of Saccharomyces
cerevisiae appears to produce a defective delta-9 fatty acid desaturase.
This enzyme catalyzes double bond formation between carbons 9 and 10 of
palmitoyl and stearoyl coenzyme A. A DNA fragment isolated by
complementation of an ole1 strain repairs the ufa requirement in mutant
cells. Genetic analysis of the cloned DNA fragment indicates that it is
allelic to the OLE1 gene. Disruption of a single copy of the wild type gene
in a diploid strain produces both wild type and nonreverting ufa-requiring
haploid progeny upon sporulation. Membrane lipids of the disrupted haploid
strains contain only ufas supplied in the growth medium. The recovery of
activity in both wild type and disrupted segregants was examined after
removal of ufas from the growth medium. Following ufa deprivation
disruptant cells grew normally for about three generations and then at a
slower rate for at least 0.6 generations. During that time cellular ufas
dropped from 63 to 7.3 mol % of the total fatty acids. No production of the
16:1 and 18:1 products of the desaturase was observed in disruptant cells,
whereas desaturation in wild type control cells was evident 2 h after
deprivation. These results indicate that 1) the OLE1 gene is essential for
production of monounsaturated fatty acids and is probably the structural
gene for the delta-9 desaturase enzyme. 2) A large part of membrane ufas
present under normal culture conditions are not essential for growth and
cell division.
Isolation and characterization of OLE1, a gene affecting fatty acid desaturation from Saccharomyces cerevisiae
Nelson Biological Laboratory, Bureau of Biological Research, Rutgers University, Piscataway, New Jersey 08855-1059.
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