Saccharomyces cerevisiae MATa mutant cells defective in pointed projection formation in response to alpha-factor at...

We have isolated Saccharomyces cerevisiae MATa mutant cells that do not form a pointed projection but elongate in response to alpha-factor at high concentrations. Complementation tests defined three genes, PPF1, PPF2, and PPF3 (for pointed projection formation), necessary for pointed projection formation. Allelism tests with genes known to be needed for projection formation revealed that PPF1 is identical to SPA2, while PPF2 and PPF3 are not allelic to SST2, STE2, SPA2, BEM1 or SLK1/SSP31/BCK1. The morphology of MATa ppf mutants treated with high concentrations of alpha-factor is similar to that of MATa PPF cells treated with alpha-factor at low concentrations. Quantitative mating tests showed that PPF2 and PPF3 are not essential for mating in either MATa or MAT alpha background. Monitoring of division arrest and expression of an alpha-factor-inducible gene revealed that mutations in the PPF genes do not affect the responses of MATa cells to low concentrations of alpha-factor. Unlike wild-type cells, the ppf mutants exhibited early recovery from alpha-factor-induced division arrest. Furthermore, vegetatively growing ppf3-1 cells are slightly defective in cell separation of mother and daughter cells and in selection of the correct bud sites in all cell types. These results indicate that PPF2 and PPF3 are involved in the response to alpha-factor at high concentrations and that PPF3 is also required for proper establishment of polarity in vegetative growth.