SEC3 mutations are synthetically lethal with profilin mutations and cause defects in diploid-specific bud-site selection.
Replacement of the wild-type yeast profilin gene (PFY1) with a mutated form (pfy1-111) that has codon 72 changed to encode glutamate rather than arginine results in defects similar to, but less severe than, those that result from complete deletion of the profilin gene. We have used a colony color-sectoring assay to identify mutations that cause pfy1-111, but not wild-type, cells to be inviable. These profilin synthetic lethal (psl) mutations result in various degrees of abnormal growth, morphology, and temperature sensitivity in PFY1 cells. We have examined psl1 strains in the most detail. Interestingly, these strains display a diploid-specific defect in bud-site selection; haploid strains bud normally, while homozygous diploid strains show a dramatic increase in random budding. We discovered that PSL1 is the late secretory gene, SEC3, and have found that mutations in several other late secretory genes are also synthetically lethal with pfy1-111. Our results are likely to reflect an interdependence between the actin cytoskeleton and secretory processes in directing cell polarity and growth. Moreover, they indicate that the secretory pathway is especially crucial for maintaining budding polarity in diploids.
Haarer
B K
BK
Department of Anatomy and Cell Biology, University of Michigan, Ann Arbor 48109, USA. haarer@mail.utexas.edu
Corbett
A
A
Kweon
Y
Y
Petzold
A S
AS
Silver
P
P
Brown
S S
SS
D000199:Actins; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003285:Contractile Proteins; D004271:DNA, Fungal; D004171:Diploidy; D005656:Fungal Proteins / Q000235:genetics*; D005804:Genes, Lethal; D008840:Microfilament Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D009994:Osmolar Concentration; D010641:Phenotype; D051304:Profilins; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D017422:Sequence Analysis, DNA; D017384:Sequence Deletion
D000199:Actins; D003285:Contractile Proteins; D004271:DNA, Fungal; D005656:Fungal Proteins; D008840:Microfilament Proteins; D051304:Profilins; C103130:SEC3 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins