A and alpha supernatant pretreatment of Saccharomyces cerevisiae cells affects both the kinetics and efficiency of mating

Abstract

The effects of culture supernatant treatment on subsequent matings between pretreated a and alpha Saccharomyces cerevisiae cells were studied. For each experiment, pairs of a and alpha [rho+] or [rho- rho0] cells in the logarithmic growth phase in defined minimal medium were pretreated for a total of 15 min (by exchanging their cell-free supernatants or by mixing samples of a and alpha cell cultures) and then mated in defined minimal (YNB) or enriched (YEP) liquid medium. All pretreated cells, regardless of treatment procedure, initiated cell fusion 15 to 35 min faster than did their nontreated counterparts. In all cases, pretreated cells mated 8 to 20% more efficiently than did nonpretreated ones. Regardless of the strains, the hierarchy of mating efficiency was always treated YEP greater than untreated YEP greater than treated YNB greater than untreated YNB. The cell fusion kinetics in alpha [rho+] X a [rho-] crosses were most affected by pretreatment (delta 30 to 35 min), whereas [rho+] X [rho+] crosses were least affected (delta 15 min). These results are discussed in relation to the functions known for a and alpha pheromones. The successful pretreatment regimes were used to design new rapid and efficient techniques for mating YNB-grown log-phase cells in either YNB or YEP liquid media. These techniques can be used for small- or large-scale mating, and because of their inherent media flexibility, they have many potential applications to future studies on mating-specific or intrazygotic phenomena.