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Published 12 November 2001. doi:10.1083/jcb.200106065
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© The Rockefeller University Press, 0021-9525/2001/11/581 $5.00
The Journal of Cell Biology, Volume 155, Number 4, November 12, 2001 581-592

Article

 Yeast Cdc42 functions at a late step in exocytosis, specifically during polarized growth of the emerging bud

Joan E. Adamo1,2, John J. Moskow3, Amy S. Gladfelter3, Domenic Viterbo2, Daniel J. Lew3 and Patrick J. Brennwald1,2

1 Department of Cell and Developmental Biology, University of North Carolina at Chapel Hill, Chapel Hill, NC 27599
2 Department of Cell Biology, Weill Medical College of Cornell University, New York, NY 10021
3 Department of Pharmacology and Cancer Biology, Duke University Medical Center, Durham, NC 27710

Address correspondence to Patrick J. Brennwald, Dept. of Cell and Developmental Biology, University of North Carolina at Chapel Hill, 536 Taylor Hall, CB #7090, Chapel Hill, NC 27599-7090. Tel.: (919) 843-4995. Fax: (919) 966-1856. E-mail: pjbrennw{at}med.unc.edu

The Rho family GTPase Cdc42 is a key regulator of cell polarity and cytoskeletal organization in eukaryotic cells. In yeast, the role of Cdc42 in polarization of cell growth includes polarization of the actin cytoskeleton, which delivers secretory vesicles to growth sites at the plasma membrane. We now describe a novel temperature-sensitive mutant, cdc42-6, that reveals a role for Cdc42 in docking and fusion of secretory vesicles that is independent of its role in actin polarization. cdc42-6 mutants can polarize actin and deliver secretory vesicles to the bud, but fail to fuse those vesicles with the plasma membrane. This defect is manifested only during the early stages of bud formation when growth is most highly polarized, and appears to reflect a requirement for Cdc42 to maintain maximally active exocytic machinery at sites of high vesicle throughput. Extensive genetic interactions between cdc42-6 and mutations in exocytic components support this hypothesis, and indicate a functional overlap with Rho3, which also regulates both actin organization and exocytosis. Localization data suggest that the defect in cdc42-6 cells is not at the level of the localization of the exocytic apparatus. Rather, we suggest that Cdc42 acts as an allosteric regulator of the vesicle docking and fusion apparatus to provide maximal function at sites of polarized growth.

Key Words: Cdc42; Rho; GTPases; exocytosis; cell polarity


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