Actin and septin ultrastructures at the budding yeast cell cortex.

Budding yeast has been a powerful model organism for studies of the roles of actin in endocytosis and septins in cell division and in signaling. However, the depth of mechanistic understanding that can be obtained from such studies has been severely hindered by a lack of ultrastructural information about how actin and septins are organized at the cell cortex. To address this problem, we developed rapid-freeze and deep-etch techniques to image the yeast cell cortex in spheroplasted cells at high resolution. The cortical actin cytoskeleton assembles into conical or mound-like structures composed of short, cross-linked filaments. The Arp2/3 complex localizes near the apex of these structures, suggesting that actin patch assembly may be initiated from the apex. Mutants in cortical actin patch components with defined defects in endocytosis disrupted different stages of cortical actin patch assembly. Based on these results, we propose a model for actin function during endocytosis. In addition to actin structures, we found that septin-containing filaments assemble into two kinds of higher order structures at the cell cortex: rings and ordered gauzes. These images provide the first high-resolution views of septin organization in cells.

[1]  J. Thorner,et al.  Protein-protein interactions governing septin heteropentamer assembly and septin filament organization in Saccharomyces cerevisiae. , 2004, Molecular biology of the cell.

[2]  J. Cooper,et al.  Yeast actin patches are networks of branched actin filaments , 2004, The Journal of cell biology.

[3]  Yves Barral,et al.  Spatial Coordination of Cytokinetic Events by Compartmentalization of the Cell Cortex , 2004, Science.

[4]  Å. Engqvist-Goldstein,et al.  RNAi-mediated Hip1R silencing results in stable association between the endocytic machinery and the actin assembly machinery. , 2004, Molecular biology of the cell.

[5]  Å. Engqvist-Goldstein,et al.  Actin assembly and endocytosis: from yeast to mammals. , 2003, Annual review of cell and developmental biology.

[6]  David G. Drubin,et al.  A Pathway for Association of Receptors, Adaptors, and Actin during Endocytic Internalization , 2003, Cell.

[7]  J. Caviston,et al.  The role of Cdc42p GTPase-activating proteins in assembly of the septin ring in yeast. , 2003, Molecular biology of the cell.

[8]  Chao Zhang,et al.  Dynamic phosphoregulation of the cortical actin cytoskeleton and endocytic machinery revealed by real-time chemical genetic analysis , 2003, The Journal of cell biology.

[9]  M. Longtine,et al.  Regulation of septin organization and function in yeast. , 2003, Trends in cell biology.

[10]  Anya L. Goodman,et al.  The Saccharomyces cerevisiae calponin/transgelin homolog Scp1 functions with fimbrin to regulate stability and organization of the actin cytoskeleton. , 2003, Molecular biology of the cell.

[11]  A. Rodal,et al.  Negative Regulation of Yeast WASp by Two SH3 Domain-Containing Proteins , 2003, Current Biology.

[12]  K. Ayscough,et al.  An interaction between Sla1p and Sla2p plays a role in regulating actin dynamics and endocytosis in budding yeast , 2003, Journal of Cell Science.

[13]  S. Ishijima,et al.  Ultrastructure and behavior of actin cytoskeleton during cell wall formation in the fission yeast Schizosaccharomyces pombe. , 2003, Journal of electron microscopy.

[14]  Matthew S. Gentry,et al.  Phosphorylation-dependent regulation of septin dynamics during the cell cycle. , 2003, Developmental cell.

[15]  A. Bretscher,et al.  Microfilaments and microtubules: the news from yeast. , 2002, Current opinion in microbiology.

[16]  A. Straight,et al.  Self- and actin-templated assembly of Mammalian septins. , 2002, Developmental cell.

[17]  Y. Barral,et al.  Septins: a ring to part mother and daughter , 2002, Current Genetics.

[18]  P. D. Andrews,et al.  Sla1p couples the yeast endocytic machinery to proteins regulating actin dynamics. , 2002, Journal of cell science.

[19]  G. Payne,et al.  Sla1p serves as the targeting signal recognition factor for NPFX(1,2)D-mediated endocytosis , 2002, The Journal of cell biology.

[20]  K. Takeo,et al.  Electron microscopy of pathogenic yeasts Cryptococcus neoformans and Exophiala dermatitidis by high-pressure freezing. , 2002, Journal of electron microscopy.

[21]  A. Rodal,et al.  Modular complexes that regulate actin assembly in budding yeast. , 2001, Current opinion in microbiology.

[22]  B. Wendland,et al.  Clathrin Function in Yeast Endocytosis , 2001, Traffic.

[23]  H. Pelham SNAREs and the specificity of membrane fusion. , 2001, Trends in cell biology.

[24]  R. Pelham,et al.  Role of actin polymerization and actin cables in actin-patch movement in Schizosaccharomyces pombe , 2001, Nature Cell Biology.

[25]  M. Gabriel,et al.  Microtubules and actin cytoskeleton in Cryptococcus neoformans compared with ascomycetous budding and fission yeasts. , 2001, European journal of cell biology.

[26]  A. Munn The yeast endocytic membrane transport system , 2000, Microscopy research and technique.

[27]  J. Derisi,et al.  Plasma membrane compartmentalization in yeast by messenger RNA transport and a septin diffusion barrier. , 2000, Science.

[28]  P. Silver,et al.  Mutants Affecting the Structure of the Cortical Endoplasmic Reticulum in Saccharomyces cerevisiae , 2000, The Journal of cell biology.

[29]  J. Heuser The Production of ‘Cell Cortices’ for Light and Electron Microscopy , 2000, Traffic.

[30]  M. Snyder,et al.  Compartmentalization of the cell cortex by septins is required for maintenance of cell polarity in yeast. , 2000, Molecular cell.

[31]  A. Svoboda,et al.  Cell wall and cytoskeleton reorganization as the response to hyperosmotic shock in Saccharomyces cerevisiae , 2000, Archives of Microbiology.

[32]  A. Bretscher,et al.  Polarization of cell growth in yeast. , 2000, Journal of cell science.

[33]  J. Heuser Membrane Traffic in Anaglyph Stereo , 2000, Traffic.

[34]  B. Kay,et al.  The EH network. , 1999, Experimental cell research.

[35]  J. McIntosh,et al.  High-voltage electron tomography of spindle pole bodies and early mitotic spindles in the yeast Saccharomyces cerevisiae. , 1999, Molecular biology of the cell.

[36]  Pietro De Camilli,et al.  Functional partnership between amphiphysin and dynamin in clathrin-mediated endocytosis , 1999, Nature Cell Biology.

[37]  J. Yates,et al.  Coronin Promotes the Rapid Assembly and Cross-linking of Actin Filaments and May Link the Actin and Microtubule Cytoskeletons in Yeast , 1999, The Journal of cell biology.

[38]  M. Mann,et al.  Polymerization of Purified Yeast Septins: Evidence That Organized Filament Arrays May Not Be Required for Septin Function , 1998, The Journal of cell biology.

[39]  M. Longtine,et al.  Role of the Yeast Gin4p Protein Kinase in Septin Assembly and the Relationship between Septin Assembly and Septin Function , 1998, The Journal of cell biology.

[40]  James G. McNally,et al.  Assembly and Function of the Actin Cytoskeleton of Yeast: Relationships between Cables and Patches , 1998, The Journal of cell biology.

[41]  T D Pollard,et al.  The interaction of Arp2/3 complex with actin: nucleation, high affinity pointed end capping, and formation of branching networks of filaments. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[42]  Y. Takai,et al.  Isolation and characterization of a novel actin filament-binding protein from Saccharomyces cerevisiae , 1998, Oncogene.

[43]  M. Mann,et al.  The complex containing actin-related proteins Arp2 and Arp3 is required for the motility and integrity of yeast actin patches , 1997, Current Biology.

[44]  A. Hirata,et al.  Vam3p, a new member of syntaxin related protein, is required for vacuolar assembly in the yeast Saccharomyces cerevisiae. , 1997, Journal of cell science.

[45]  D. Drubin,et al.  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 , 1997, The Journal of cell biology.

[46]  B. Dujon,et al.  The Saccharomyces cerevisiae actin-related protein Arp2 is involved in the actin cytoskeleton , 1996, The Journal of cell biology.

[47]  H. Warrick,et al.  Synthetic lethality screen identifies a novel yeast myosin I gene (MYO5): myosin I proteins are required for polarization of the actin cytoskeleton , 1996, The Journal of cell biology.

[48]  D. Botstein,et al.  Movement of yeast cortical actin cytoskeleton visualized in vivo. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[49]  H. Riezman,et al.  Role of Type I Myosins in Receptor-Mediated Endocytosis in Yeast , 1996, Science.

[50]  J. Cooper,et al.  Movement of cortical actin patches in yeast , 1996, The Journal of cell biology.

[51]  Brian J. Stevenson,et al.  end5, end6, and end7: mutations that cause actin delocalization and block the internalization step of endocytosis in Saccharomyces cerevisiae. , 1995, Molecular biology of the cell.

[52]  M. Gabriel,et al.  Actin cortical cytoskeleton and cell wall synthesis in regenerating protoplasts of the Saccharomyces cerevisiae actin mutant DBY 1693. , 1995, Microbiology.

[53]  J. Haber,et al.  Dominant lethal mutations in the plasma membrane H(+)-ATPase gene of Saccharomyces cerevisiae. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[54]  D. Botstein,et al.  Ultrastructure of the yeast actin cytoskeleton and its association with the plasma membrane , 1994, The Journal of cell biology.

[55]  D. Drubin,et al.  Synthetic-lethal interactions identify two novel genes, SLA1 and SLA2, that control membrane cytoskeleton assembly in Saccharomyces cerevisiae , 1993, The Journal of cell biology.

[56]  H. Riezman,et al.  Actin and fimbrin are required for the internalization step of endocytosis in yeast. , 1993, The EMBO journal.

[57]  J. Rohrer,et al.  end3 and end4: two mutants defective in receptor-mediated and fluid- phase endocytosis in Saccharomyces cerevisiae , 1993, The Journal of cell biology.

[58]  K. Tanaka,et al.  Dynamics of cytoplasmic organelles in the cell cycle of the fission yeast Schizosaccharomyces pombe: three-dimensional reconstruction from serial sections. , 1989, Journal of cell science.

[59]  D. Botstein,et al.  A yeast actin-binding protein is encoded by SAC6, a gene found by suppression of an actin mutation. , 1989, Science.

[60]  D Botstein,et al.  Yeast actin-binding proteins: evidence for a role in morphogenesis , 1988, The Journal of cell biology.

[61]  B. Haarer,et al.  Immunofluorescence localization of the Saccharomyces cerevisiae CDC12 gene product to the vicinity of the 10-nm filaments in the mother-bud neck , 1987, Molecular and cellular biology.

[62]  J. Pringle,et al.  Relationship of actin and tubulin distribution to bud growth in wild- type and morphogenetic-mutant Saccharomyces cerevisiae , 1984, The Journal of cell biology.

[63]  B. Byers,et al.  A highly ordered ring of membrane-associated filaments in budding yeast , 1976, The Journal of cell biology.

[64]  D. Demarini,et al.  A Bni4-Glc7 phosphatase complex that recruits chitin synthase to the site of bud emergence. , 2003, Molecular biology of the cell.

[65]  B. Goode,et al.  Purification of yeast actin and actin-associated proteins. , 2002, Methods in enzymology.

[66]  R. Mullins,et al.  Cellular control of actin nucleation. , 2002, Annual review of cell and developmental biology.

[67]  H. Riezman,et al.  Morphology of the yeast endocytic pathway. , 1998, Molecular biology of the cell.

[68]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[69]  of the S. , 2022 .