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© The Rockefeller University Press, 0021-9525/1999//845 $5.00
The Journal of Cell Biology, Volume 147, Number 4, , 1999 845-856

Original Article

Pak-Family Kinases Regulate Cell and Actin Polarization Throughout the Cell Cycle of Saccharomyces cerevisiae



Stephen P. Hollya and Kendall J. Blumera

a Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, Missouri 63110
Department of Cell Biology and Physiology, Washington University School of Medicine, 660 S. Euclid Ave., Box 8228, St. Louis, MO 63110.(314) 362-7463(314) 362-1668

kblumer{at}cellbio.wustl.edu

During the cell cycle of the yeast Saccharomyces cerevisiae, the actin cytoskeleton and cell surface growth are polarized, mediating bud emergence, bud growth, and cytokinesis. We have determined whether p21-activated kinase (PAK)-family kinases regulate cell and actin polarization at one or several points during the yeast cell cycle. Inactivation of the PAK homologues Ste20 and Cla4 at various points in the cell cycle resulted in loss of cell and actin cytoskeletal polarity, but not in depolymerization of F-actin. Loss of PAK function in G1 depolarized the cortical actin cytoskeleton and blocked bud emergence, but allowed isotropic growth and led to defects in septin assembly, indicating that PAKs are effectors of the Rho–guanosine triphosphatase Cdc42. PAK inactivation in S/G2 resulted in depolarized growth of the mother and bud and a loss of actin polarity. Loss of PAK function in mitosis caused a defect in cytokinesis and a failure to polarize the cortical actin cytoskeleton to the mother-bud neck. Cla4–green fluorescent protein localized to sites where the cortical actin cytoskeleton and cell surface growth are polarized, independently of an intact actin cytoskeleton. Thus, PAK family kinases are primary regulators of cell and actin cytoskeletal polarity throughout most or all of the yeast cell cycle. PAK-family kinases in higher organisms may have similar functions.

Key Words: p21-activated kinase • actin • polarity • cell cycle • Cdc42

© 1999 The Rockefeller University Press

1.used in this paper: DAPI, 4,6-diamidino-2-phenylindole; DHFR, dihydrofolate reductase; GFP, green fluorescent protein; HA, hemagglutinin; ORF, open reading frame; PAK, p21-activated kinase; YPD, rich media containing yeast extract, bactopeptone, and dextrose


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