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© The Rockefeller University Press, 0021-9525/1998//461 $5.00
The Journal of Cell Biology, Volume 140, Number 3, , 1998 461-483

Article

Pheromone-regulated Genes Required for Yeast Mating Differentiation



Scott Erdman, Li Lin, Michael Malczynski, and Michael Snyder

Department of Biology, Yale University, New Haven, Connecticut 06520-8103

Yeast cells mate by an inducible pathway that involves agglutination, mating projection formation, cell fusion, and nuclear fusion. To obtain insight into the mating differentiation of Saccharomyces cerevisiae, we carried out a large-scale transposon tagging screen to identify genes whose expression is regulated by mating pheromone. 91,200 transformants containing random lacZ insertions were screened for β-galactosidase (β-gal) expression in the presence and absence of {alpha} factor, and 189 strains containing pheromone-regulated lacZ insertions were identified. Transposon insertion alleles corresponding to 20 genes that are novel or had not previously been known to be pheromone regulated were examined for effects on the mating process. Mutations in four novel genes, FIG1, FIG2, KAR5/ FIG3, and FIG4 were found to cause mating defects. Three of the proteins encoded by these genes, Fig1p, Fig2p, and Fig4p, are dispensible for cell polarization in uniform concentrations of mating pheromone, but are required for normal cell polarization in mating mixtures, conditions that involve cell–cell communication. Fig1p and Fig2p are also important for cell fusion and conjugation bridge shape, respectively. The fourth protein, Kar5p/Fig3p, is required for nuclear fusion. Fig1p and Fig2p are likely to act at the cell surface as Fig1:: β-gal and Fig2::β-gal fusion proteins localize to the periphery of mating cells. Fig4p is a member of a family of eukaryotic proteins that contain a domain homologous to the yeast Sac1p. Our results indicate that a variety of novel genes are expressed specifically during mating differentiation to mediate proper cell morphogenesis, cell fusion, and other steps of the mating process.

Abbreviations used in this paper: β-gal, β-galactosidase; DAPI, 4',6-diamidino-Z-phenylindole; DIC, differential interference-contrast microscopy; FIG, factor-induced gene; MAP, mitogen-activated kinase; ORF, open reading frame; PEG, polyethylene glycol; PRE, pheromone response element; SC, synthetic complete medium; TM, transmembrane; TMD, transmembrane domain; YPD, yeast extract/peptone/dextrose medium.

S. Erdman was supported by an American Cancer Society Postdoctoral Fellowship. This research was supported by National Institutes of Health grants (HD32637 and GM36494).

Address all correspondence to Michael Snyder, Dept. of Biology, PO Box 208103, Yale University, New Haven, CT 06520-8103. Tel.: (203) 432-6139. Fax: (203) 432-6161. E-mail: michael.snyder{at}yale.edu


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