The Journal of Cell Biology, Vol 128, 599-615, Copyright © 1995 by The Rockefeller University Press

ARTICLES

Regulation of cortical actin cytoskeleton assembly during polarized cell growth in budding yeast

R Li, Y Zheng and DG Drubin
Department of Molecular and Cell Biology, University of California, Berkeley 94720.

We have established an in vitro assay for assembly of the cortical actin cytoskeleton of budding yeast cells. After permeabilization of yeast by a novel procedure designed to maintain the spatial organization of cellular constituents, exogenously added fluorescently labeled actin monomers assemble into distinct structures in a pattern that is similar to the cortical actin distribution in vivo. Actin assembly in the bud of small-budded cells requires a nucleation activity provided by protein factors that appear to be distinct from the barbed ends of endogenous actin filaments. This nucleation activity is lost in cells that lack either Sla1 or Sla2, proteins previously implicated in cortical actin cytoskeleton function, suggesting a possible role for these proteins in the nucleation reaction. The rate and the extent of actin assembly in the bud are increased in permeabilized delta cap2 cells, providing evidence that capping protein regulates the ability of the barbed ends of actin filaments to grow in yeast cells. Actin incorporation in the bud can be stimulated by treating the permeabilized cells with GTP-gamma S, and, significantly, the stimulatory effect is eliminated by a mutation in CDC42, a gene that encodes a Rho-like GTP-binding protein required for bud formation. Furthermore, the lack of actin nucleation activity in the cdc42 mutant can be complemented in vitro by a constitutively active Cdc42 protein. These results suggest that Cdc42 is closely involved in regulating actin assembly during polarized cell growth.
CiteULike    Complore    Connotea    Del.icio.us    Digg    Facebook    Reddit    Technorati    Twitter    What's this?

This article has been cited by other articles:

• Park, H.-O., Bi, E. (2007). Central Roles of Small GTPases in the Development of Cell Polarity in Yeast and Beyond. Microbiol. Mol. Biol. Rev. 71: 48-96 [Abstract] [Full Text]  
• Kovar, D. R., Wu, J.-Q., Pollard, T. D. (2005). Profilin-mediated Competition between Capping Protein and Formin Cdc12p during Cytokinesis in Fission Yeast. Mol. Biol. Cell 16: 2313-2324 [Abstract] [Full Text]  
• Young, M. E., Cooper, J. A., Bridgman, P. C. (2004). Yeast actin patches are networks of branched actin filaments. JCB 166: 629-635 [Abstract] [Full Text]  
• Kim, K., Yamashita, A., Wear, M. A., Maeda, Y., Cooper, J. A. (2004). Capping protein binding to actin in yeast: biochemical mechanism and physiological relevance. JCB 164: 567-580 [Abstract] [Full Text]  
• Goehring, A. S., Rivers, D. M., Sprague, G. F. Jr. (2003). Urmylation: A Ubiquitin-like Pathway that Functions during Invasive Growth and Budding in Yeast. Mol. Biol. Cell 14: 4329-4341 [Abstract] [Full Text]  
• Carnahan, R. H., Gould, K. L. (2003). The PCH family protein, Cdc15p, recruits two F-actin nucleation pathways to coordinate cytokinetic actin ring formation in Schizosaccharomyces pombe. JCB 162: 851-862 [Abstract] [Full Text]  
• Goehring, A. S., Mitchell, D. A., Tong, A. H. Y., Keniry, M. E., Boone, C., Sprague, G. F. Jr. (2003). Synthetic Lethal Analysis Implicates Ste20p, a p21-activated Protein Kinase, in Polarisome Activation. Mol. Biol. Cell 14: 1501-1516 [Abstract] [Full Text]  
• Hannus, M., Feiguin, F., Heisenberg, C.-P., Eaton, S. (2003). Planar cell polarization requires Widerborst, a B' regulatory subunit of protein phosphatase 2A. Development 129: 3493-3503 [Abstract] [Full Text]  
• Kaminska, J., Gajewska, B., Hopper, A. K., Zoladek, T. (2002). Rsp5p, a New Link between the Actin Cytoskeleton and Endocytosis in the Yeast Saccharomyces cerevisiae. Mol. Cell. Biol. 22: 6946-6948 [Abstract] [Full Text]  
• Drees, B. L., Sundin, B., Brazeau, E., Caviston, J. P., Chen, G.-C., Guo, W., Kozminski, K. G., Lau, M. W., Moskow, J. J., Tong, A., Schenkman, L. R., McKenzie, A. III, Brennwald, P., Longtine, M., Bi, E., Chan, C., Novick, P., Boone, C., Pringle, J. R., Davis, T. N., Fields, S., Drubin, D. G. (2001). A protein interaction map for cell polarity development. JCB 154: 549-576 [Abstract] [Full Text]  
• Gladfelter, A. S., Moskow, J. J., Zyla, T. R., Lew, D. J. (2001). Isolation and Characterization of Effector-Loop Mutants of CDC42 in Yeast. Mol. Biol. Cell 12: 1239-1255 [Abstract] [Full Text]  
• Asleson, C. M., Bensen, E. S., Gale, C. A., Melms, A.-S., Kurischko, C., Berman, J. (2001). Candida albicans INT1-Induced Filamentation in Saccharomyces cerevisiae Depends on Sla2p. Mol. Cell. Biol. 21: 1272-1284 [Abstract] [Full Text]  
• Smith, M., Swamy, S., Pon, L. (2001). The life cycle of actin patches in mating yeast. J. Cell Sci. 114: 1505-1513 [Abstract]  
• Chung, C. Y., Lee, S., Briscoe, C., Ellsworth, C., Firtel, R. A. (2000). Role of Rac in controlling the actin cytoskeleton and chemotaxis in motile cells. Proc. Natl. Acad. Sci. USA 97: 5225-5230 [Abstract] [Full Text]  
• Hurtado, C. A. R., Beckerich, J.-M., Gaillardin, C., Rachubinski, R. A. (2000). A Rac Homolog Is Required for Induction of Hyphal Growth in the Dimorphic Yeast Yarrowia lipolytica. J. Bacteriol. 182: 2376-2386 [Abstract] [Full Text]  
• Bi, E., Chiavetta, J. B., Chen, H., Chen, G.-C., Chan, C. S. M., Pringle, J. R. (2000). Identification of Novel, Evolutionarily Conserved Cdc42p-interacting Proteins and of Redundant Pathways Linking Cdc24p and Cdc42p to Actin Polarization in Yeast. Mol. Biol. Cell 11: 773-793 [Abstract] [Full Text]  
• Lechler, T., Shevchenko, A., Shevchenko, A., Li, R. (2000). Direct Involvement of Yeast Type I Myosins in Cdc42-Dependent Actin Polymerization. JCB 148: 363-374 [Abstract] [Full Text]  
• Holly, S. P., Blumer, K. J. (1999). Pak-Family Kinases Regulate Cell and Actin Polarization Throughout the Cell Cycle of Saccharomyces cerevisiae. JCB 147: 845-856 [Abstract] [Full Text]  
• Chung, C. Y., Firtel, R. A. (1999). Paka, a Putative Pak Family Member, Is Required for Cytokinesis and the Regulation of the Cytoskeleton in Dictyostelium discoideum Cells during Chemotaxis. JCB 147: 559-576 [Abstract] [Full Text]  
• Delley, P.-A., Hall, M. N. (1999). Cell Wall Stress Depolarizes Cell Growth via Hyperactivation of Rho1. JCB 147: 163-174 [Abstract] [Full Text]  
• Bailleul, P. A., Newnam, G. P., Steenbergen, J. N., Chernoff, Y. O. (1999). Genetic Study of Interactions Between the Cytoskeletal Assembly Protein Sla1 and Prion-Forming Domain of the Release Factor Sup35 (eRF3) in Saccharomyces cerevisiae. Genetics 153: 81-94 [Abstract] [Full Text]  
• Yang, S., Cope, M. J. T. V., Drubin, D. G. (1999). Sla2p Is Associated with the Yeast Cortical Actin Cytoskeleton via Redundant Localization Signals. Mol. Biol. Cell 10: 2265-2283 [Abstract] [Full Text]  
• Lau, P.-m., Zucker, R. S., Bentley, D. (1999). Induction of Filopodia by Direct Local Elevation of Intracellular Calcium Ion Concentration. JCB 145: 1265-1276 [Abstract] [Full Text]  
• Robinson, N. G. G., Guo, L., Imai, J., Toh-e, A., Matsui, Y., Tamanoi, F. (1999). Rho3 of Saccharomyces cerevisiae, Which Regulates the Actin Cytoskeleton and Exocytosis, Is a GTPase Which Interacts with Myo2 and Exo70. Mol. Cell. Biol. 19: 3580-3587 [Abstract] [Full Text]  
• Ayscough, K. R., Eby, J. J., Lila, T., Dewar, H., Kozminski, K. G., Drubin, D. G. (1999). Sla1p Is a Functionally Modular Component of the Yeast Cortical Actin Cytoskeleton Required for Correct Localization of Both Rho1p-GTPase and Sla2p, a Protein with Talin Homology. Mol. Biol. Cell 10: 1061-1075 [Abstract] [Full Text]  
• Cope, M.J. T.V., Yang, S., Shang, C., Drubin, D. G. (1999). Novel Protein Kinases Ark1p and Prk1p Associate with and Regulate the Cortical Actin Cytoskeleton in Budding Yeast. JCB 144: 1203-1218 [Abstract] [Full Text]  
• Johnson, D. I. (1999). Cdc42: An Essential Rho-Type GTPase Controlling Eukaryotic Cell Polarity. Microbiol. Mol. Biol. Rev. 63: 54-105 [Abstract] [Full Text]  
• Blader, I. J., Cope, M. J. T. V., Jackson, T. R., Profit, A. A., Greenwood, A. F., Drubin, D. G., Prestwich, G. D., Theibert, A. B. (1999). GCS1, an Arf Guanosine Triphosphatase-activating Protein in Saccharomyces cerevisiae, Is Required for Normal Actin Cytoskeletal Organization In Vivo and Stimulates Actin Polymerization In Vitro. Mol. Biol. Cell 10: 581-596 [Abstract] [Full Text]  
• Shannon, K. B., Li, R. (1999). The Multiple Roles of Cyk1p in the Assembly and Function of the Actomyosin Ring in Budding Yeast. Mol. Biol. Cell 10: 283-296 [Abstract] [Full Text]  
• Zeng, G., Cai, M. (1999). Regulation of the Actin Cytoskeleton Organization in Yeast by a Novel Serine/Threonine Kinase Prk1p. JCB 144: 71-82 [Abstract] [Full Text]  
• Gustin, M. C., Albertyn, J., Alexander, M., Davenport, K. (1998). MAP Kinase Pathways in the Yeast Saccharomyces cerevisiae. Microbiol. Mol. Biol. Rev. 62: 1264-1300 [Abstract] [Full Text]  
• Finger, F. P., Novick, P. (1998). Spatial Regulation of Exocytosis: Lessons from Yeast. JCB 142: 609-612 [Full Text]  
• Chen, G.-C., Kim, Y.-J., Chan, C. S.M. (1997). The Cdc42 GTPase-associated proteins Gic1 and Gic2 are required for polarized cell growth in Saccharomyces cerevisiae. Genes Dev. 11: 2958-2971 [Abstract] [Full Text]  
• Wesp, A., Hicke, L., Palecek, J., Lombardi, R., Aust, T., Munn, A.L., Riezman, H. (1997). End4p/Sla2p Interacts with Actin-associated Proteins for Endocytosis in Saccharomyces cerevisiae. Mol. Biol. Cell 8: 2291-2306 [Abstract] [Full Text]  
• Manning, B. D., Padmanabha, R., Snyder, M. (1997). The Rho-GEF Rom2p Localizes to Sites of Polarized Cell Growth and Participates in Cytoskeletal Functions in Saccharomyces cerevisiae. Mol. Biol. Cell 8: 1829-1844 [Abstract] [Full Text]  
• Mackay, D. J.G., Esch, F., Furthmayr, H., Hall, A. (1997). Rho- and Rac-dependent Assembly of Focal Adhesion Complexes and Actin Filaments in Permeabilized Fibroblasts: An Essential Role for Ezrin/Radixin/Moesin Proteins. JCB 138: 927-938 [Abstract] [Full Text]  
• Zigmond, S. H., Joyce, M., Borleis, J., Bokoch, G. M., Devreotes, P. N. (1997). Regulation of Actin Polymerization in Cell-free Systems by GTP{gamma}S and Cdc42. JCB 138: 363-374 [Abstract] [Full Text]  
• Lechler, T., Li, R. (1997). In Vitro Reconstitution of Cortical Actin Assembly Sites in Budding Yeast. JCB 138: 95-103 [Abstract] [Full Text]  
• McCann, R. O., Craig, S. W. (1997). The I/LWEQ module: a conserved sequence that signifies F-actin binding in functionally diverse proteins from yeast to mammals. Proc. Natl. Acad. Sci. USA 94: 5679-5684 [Abstract] [Full Text]  
• Ayscough, K. R., Stryker, J., Pokala, N., Sanders, M., Crews, P., Drubin, D. G. (1997). High Rates of Actin Filament Turnover in Budding Yeast and Roles for Actin in Establishment and Maintenance of Cell Polarity Revealed Using the Actin Inhibitor Latrunculin-A. JCB 137: 399-416 [Abstract] [Full Text]  
• Evangelista, M., Blundell, K., Longtine, M. S., Chow, C. J., Adames, N., Pringle, J. R., Peter, M., Boone, C. (1997). Bni1p, a Yeast Formin Linking Cdc42p and the Actin Cytoskeleton During Polarized Morphogenesis. Science 276: 118-122 [Abstract] [Full Text]  
• Li, R. (1997). Bee1, a Yeast Protein with Homology to Wiscott-Aldrich Syndrome Protein, Is Critical for the Assembly of Cortical Actin Cytoskeleton. JCB 136: 649-658 [Abstract] [Full Text]  
• van Deurs, B, von Bulow, F, Vilhardt, F, Holm, P., Sandvig, K (1996). Destabilization of plasma membrane structure by prevention of actin polymerization. Microtubule-dependent tubulation of the plasma membrane. J. Cell Sci. 109: 1655-1665 [Abstract]  
• Pringle, J.R., Bi, E., Harkins, H.A., Zahner, J.E., De Virgilio, C., Chant, J., Corrado, K., Fares, H. (1995). Establishment of Cell Polarity in Yeast. Cold Spring Harb Symp Quant Biol 60: 729-744 [Abstract]  
• Hackam, A. S., Yassa, A. S., Singaraja, R., Metzler, M., Gutekunst, C.-A., Gan, L., Warby, S., Wellington, C. L., Vaillancourt, J., Chen, N., Gervais, F. G., Raymond, L., Nicholson, D. W., Hayden, M. R. (2000). Huntingtin Interacting Protein 1 Induces Apoptosis via a Novel Caspase-dependent Death Effector Domain. J. Biol. Chem. 275: 41299-41308 [Abstract] [Full Text]  

Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents