Komagataella phaffii YPS1-5 encodes the alpha-factor degrading protease Bar1

Yeast mating pheromones are small secreted peptides required for efficient mating between cells of opposite mating type. Pheromone gradients allow the cells to detect potential mating partners. Secreted pheromone degrading proteases steepen local gradients and allow fast recovery from the pheromone signal. The methylotrophic yeast Komagataella phaffii is a preferentially haploid species. Only under nitrogen starvation, mating genes are activated and the cells are able to undergo a full sexual cycle of mating and sporulation. It has been shown that, similar to other yeasts, K. phaffii requires the mating pheromone and pheromone surface receptor genes for efficient mating. The analysis of so far uncharacterized mating-type-specific genes allowed us to identify the K. phaffii α-factor protease gene YPS1-5. It encodes an aspartic protease of the yapsin family and is upregulated only in a-type cells under mating conditions. The phenotype of K. phaffiia-type strains with a deletion in the protease gene was found to be highly similar to the phenotype of Saccharomyces cerevisiae α-factor protease BAR1 deletion strains. They are highly sensitive to α-factor pheromone in pheromone sensitivity assays and were found to mate with reduced efficiency. Based on our results, we propose to rename the gene into K. phaffii BAR1.