Combining mutations in the incoming and outgoing pheromone signal pathways causes a synergistic mating defect in...

Mating pheromones stimulate Saccharomyces cerevisiae yeast cells to form a pointed projection that becomes the site of cell fusion during conjugation. To investigate the role of mating projections, we screened for mutations that enhanced the weak mating defect of MATa ste2-T326 cells that are defective in forming pointed projections. These cells are also 10-fold more sensitive to alpha-factor pheromone because ste2-T326 encodes truncated alpha-factor receptors that are not regulated properly. Mutations in AXL1, STE6 and FUS3 were identified in the screen. AXL1 was studied further because it is required for efficient a-factor pheromone production and for selecting the site for bud morphogenesis. Mutation of AXL1 did not enhance the morphogenesis or pheromone sensitivity defects of ste2-T326. Instead, the synergistic mating defect was apparently due to decreased a-factor production because the axl1Delta ste2-T326 cells mated well with a sst2 alpha mating partner that is supersensitive to a-factor. When combined with a wild-type mating partner, the ste2-T326 axl1Delta cells failed to mate because they did not lock cell walls, one of the earliest steps in conjugation. Analysis of axl1Delta in combination with other mutations that cause defects in morphogenesis or pheromone sensitivity (e.g. bar1, sst2, afr1) indicated that both phenotypes of ste2-T326 cells, supersensitivity to alpha-factor and the defect in forming pointed projections, contributed to the synergistic mating defect. We suggest a model that the synergistic mating defect is caused by the combined effects of ste2-T326 and axl1Delta on the presentation of a-factor to partner cells. Altogether, these results demonstrate an important linkage between the incoming and outgoing pheromone signals during the intercellular communication that promotes yeast mating.