Cell fusion during yeast mating requires high levels of a-factor mating pheromone.
During conjugation, two yeast cells fuse to form a single zygote. Cell fusion requires extensive remodeling of the cell wall, both to form a seal between the two cells and to remove the intervening material. The two plasma membranes then fuse to produce a continuous cytoplasm. We report the characterization of two cell fusion defective (Fus-) mutants, fus5 and fus8, isolated previously in our laboratory. Fluorescence and electron microscopy demonstrated that the fus5 and fus8 mutant zygotes were defective for cell wall remodeling/removal but not plasma membrane fusion. Strikingly, fus5 and fus8 were a specific; both mutations caused the mutant phenotype when present in the MATa parent but not in the MAT alpha parent. Consistent with an a-specific defect, the fus5 and fus8 mutants produced less a-factor than the isogenic wild-type strain. FUS5 and FUS8 were determined to be allelic to AXL1 and RAM1, respectively, two genes known to be required for biogenesis of a-factor. Several experiments demonstrated that the partial defect in a-factor production resulted in the Fus- phenotype. First, overexpression of a-factor in the fus mutants suppressed the Fus- defect. Second, matings to an MAT alpha partner supersensitive to mating pheromone (sst2 delta) suppressed the Fus- defect in trans. Finally, the gene encoding a-factor, MFA1, was placed under the control of a repressible promoter; reduced levels of wild-type a-factor caused an identical cell fusion defect during mating. We conclude that high levels of pheromone are required as one component of the signal for prezygotes to initiate cell fusion.
Brizzio
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V
Department of Molecular Biology, Princeton University, New Jersey 08544-1014, USA.
Gammie
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AE
Nijbroek
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G
Michaelis
S
S
Rose
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MD
D000483:Alleles; D004926:Escherichia coli / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D007339:Insulysin / Q000235:genetics; D008074:Lipoproteins / Q000235:genetics* / Q000378:metabolism*; D008666:Metalloendopeptidases; D008854:Microscopy, Electron; D008969:Molecular Sequence Data; D009154:Mutation / Q000502:physiology; D010641:Phenotype; D010675:Pheromones / Q000235:genetics / Q000378:metabolism; D010957:Plasmids; D012098:Reproduction; D012441:Saccharomyces cerevisiae / Q000502:physiology* / Q000648:ultrastructure; D029701:Saccharomyces cerevisiae Proteins; D014166:Transferases / Q000235:genetics; D015053:Zygote / Q000378:metabolism
D005656:Fungal Proteins; D008074:Lipoproteins; C095063:MFA2 protein, S cerevisiae; D010675:Pheromones; D029701:Saccharomyces cerevisiae Proteins; D014166:Transferases; C070038:RAM1 protein, S cerevisiae; C090850:AXL1 protein, S cerevisiae; D008666:Metalloendopeptidases; D007339:Insulysin