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© The Rockefeller University Press, 0021-9525/1997//111 $5.00
The Journal of Cell Biology, Volume 136, Number 1, , 1997 111-123

Article

A Role for the Actin Cytoskeleton of Saccharomyces cerevisiae in Bipolar Bud-Site Selection



Shirley Yang, Kathryn R. Ayscough, and David G. Drubin

Department of Molecular and Cell Biology, University of California, Berkeley, California 94720

Saccharomyces cerevisiae cells select bud sites according to one of two predetermined patterns. MATa and MAT{alpha} cells bud in an axial pattern, and MATa/{alpha} cells bud in a bipolar pattern. These budding patterns are thought to depend on the placement of spatial cues at specific sites in the cell cortex. Because cytoskeletal elements play a role in organizing the cytoplasm and establishing distinct plasma membrane domains, they are well suited for positioning bud-site selection cues. Indeed, the septin-containing neck filaments are crucial for establishing the axial budding pattern characteristic of MATa and MAT{alpha} cells. In this study, we determined the budding patterns of cells carrying mutations in the actin gene or in genes encoding actin-associated proteins: MATa/{alpha} cells were defective in the bipolar budding pattern, but MATa and MAT{alpha} cells still exhibit a normal axial budding pattern. We also observed that MATa/{alpha} actin cytoskeleton mutant daughter cells correctly position their first bud at the distal pole of the cell, but mother cells position their buds randomly. The actin cytoskeleton therefore functions in generation of the bipolar budding pattern and is required specifically for proper selection of bud sites in mother MATa/{alpha} cells. These observations and the results of double mutant studies support the conclusion that different rules govern bud-site selection in mother and daughter MATa/{alpha} cells. A defective bipolar budding pattern did not preclude an sla2-6 mutant from undergoing pseudohyphal growth, highlighting the central role of daughter cell bud-site selection cues in the formation of pseudohyphae. Finally, by examining the budding patterns of mad2-1 mitotic checkpoint mutants treated with benomyl to depolymerize their microtubules, we confirmed and extended previous evidence indicating that microtubules do not function in axial or bipolar bud-site selection.

Abbreviations used in this paper: mad, mitotic arrest defective; MAT, mating type; PH, pseudohyphal.

S. Yang was supported by training grants from the National Institutes of Health. K. Ayscough is an International Prize Travelling Fellow of the Wellcome Trust (038110/Z193/Z). This work was supported by grants to D.G. Drubin from the American Cancer Society (CB-106A and FRA-442).

Address all correspondence to Dr. David G. Drubin, Department of Molecular and Cell Biology, 401 Baker Hall, University of California, Berkeley, CA 94720. Tel.: (510) 642-3692 Fax: (510) 642-6420. E-mail: drubin{at}mendel.berkeley.edu


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