Actin and Endocytosis in Budding Yeast

Endocytosis, the process whereby the plasma membrane invaginates to form vesicles, is essential for bringing many substances into the cell and for membrane turnover. The mechanism driving clathrin-mediated endocytosis (CME) involves > 50 different protein components assembling at a single location on the plasma membrane in a temporally ordered and hierarchal pathway. These proteins perform precisely choreographed steps that promote receptor recognition and clustering, membrane remodeling, and force-generating actin-filament assembly and turnover to drive membrane invagination and vesicle scission. Many critical aspects of the CME mechanism are conserved from yeast to mammals and were first elucidated in yeast, demonstrating that it is a powerful system for studying endocytosis. In this review, we describe our current mechanistic understanding of each step in the process of yeast CME, and the essential roles played by actin polymerization at these sites, while providing a historical perspective of how the landscape has changed since the preceding version of the YeastBook was published 17 years ago (1997). Finally, we discuss the key unresolved issues and where future studies might be headed.

[1]  Adam C. Martin,et al.  Phosphoregulation of Arp2/3-dependent actin assembly during receptor-mediated endocytosis , 2005, Nature Cell Biology.

[2]  R. Mullins,et al.  Arp2/3 complex ATP hydrolysis promotes lamellipodial actin network disassembly but is dispensable for assembly , 2013, The Journal of cell biology.

[3]  H. Riezman,et al.  Yeast Gaa1p is required for attachment of a completed GPI anchor onto proteins , 1995, The Journal of cell biology.

[4]  Thomas D Pollard,et al.  Pathway of Actin Filament Branch Formation by Arp2/3 Complex* , 2008, Journal of Biological Chemistry.

[5]  Richard G. W. Anderson,et al.  A mutation that impairs the ability of lipoprotein receptors to localise in coated pits on the cell surface of human fibroblasts , 1977, Nature.

[6]  Alexandre V. Podtelejnikov,et al.  Mechanism of regulation of WAVE1-induced actin nucleation by Rac1 and Nck , 2002, Nature.

[7]  H. Riezman,et al.  Endocytosis is required for the growth of vacuolar H(+)-ATPase- defective yeast: identification of six new END genes , 1994, The Journal of cell biology.

[8]  Kenneth R. Henry,et al.  Protein Phosphatase-1 Binding to Scd5p Is Important for Regulation of Actin Organization and Endocytosis in Yeast* , 2002, The Journal of Biological Chemistry.

[9]  Pekka Lappalainen,et al.  Aip1p Interacts with Cofilin to Disassemble Actin Filaments , 1999, The Journal of cell biology.

[10]  D. Breitsprecher,et al.  Srv2/cyclase-associated protein forms hexameric shurikens that directly catalyze actin filament severing by cofilin , 2013, Molecular biology of the cell.

[11]  Adam C. Martin,et al.  The Journal of Cell Biology , 2002 .

[12]  X. Yu,et al.  Regulation of yeast actin cytoskeleton-regulatory complex Pan1p/Sla1p/End3p by serine/threonine kinase Prk1p. , 2001, Molecular biology of the cell.

[13]  J A McCammon,et al.  Thermodynamics and kinetics of actin filament nucleation. , 2001, Biophysical journal.

[14]  J. Cooper,et al.  Biological role and structural mechanism of twinfilin–capping protein interaction , 2004, The EMBO journal.

[15]  D. Drubin,et al.  Regulation of the Cortical Actin Cytoskeleton in Budding Yeast by Twinfilin, a Ubiquitous Actin Monomer-sequestering Protein , 1998, The Journal of cell biology.

[16]  A. Makhov,et al.  F-actin binding is essential for coronin 1B function in vivo , 2007, Journal of Cell Science.

[17]  Rebecca J Brady,et al.  Regulation of Hip1r by epsin controls the temporal and spatial coupling of actin filaments to clathrin-coated pits , 2010, Journal of Cell Science.

[18]  D. Drubin,et al.  In vivo role for actin-regulating kinases in endocytosis and yeast epsin phosphorylation. , 2001, Molecular biology of the cell.

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

[20]  J. Hurley,et al.  Syp1 is a conserved endocytic adaptor that contains domains involved in cargo selection and membrane tubulation , 2009, The EMBO journal.

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

[22]  D. Drubin,et al.  Lsb1 Is a Negative Regulator of Las17 Dependent Actin Polymerization Involved in Endocytosis , 2013, PloS one.

[23]  E. Ungewickell,et al.  Identification of the universal cofactor (auxilin 2) in clathrin coat dissociation. , 2000, European journal of cell biology.

[24]  S. Emr,et al.  A novel fluorescence-activated cell sorter-based screen for yeast endocytosis mutants identifies a yeast homologue of mammalian eps15 , 1996, The Journal of cell biology.

[25]  B. Nolen,et al.  Mechanism of a Concentration-dependent Switch between Activation and Inhibition of Arp2/3 Complex by Coronin* , 2011, The Journal of Biological Chemistry.

[26]  J. Gelles,et al.  GMF Is a Cofilin Homolog that Binds Arp2/3 Complex to Stimulate Filament Debranching and Inhibit Actin Nucleation , 2010, Current Biology.

[27]  Andrea Picco,et al.  Precise, correlated fluorescence microscopy and electron tomography of lowicryl sections using fluorescent fiducial markers. , 2012, Methods in cell biology.

[28]  Zbyszek Otwinowski,et al.  Structure and Control of the Actin Regulatory WAVE Complex , 2010, Nature.

[29]  H. Matsubara,et al.  Glia Maturation Factor-&ggr; Is Preferentially Expressed in Microvascular Endothelial and Inflammatory Cells and Modulates Actin Cytoskeleton Reorganization , 2006, Circulation research.

[30]  W. Chiu,et al.  ADF/cofilin weakens lateral contacts in the actin filament. , 1999, Journal of molecular biology.

[31]  Rong Li,et al.  Dynamics of Yeast Myosin I Evidence for a Possible Role in Scission of Endocytic Vesicles , 2004, Current Biology.

[32]  S. Ono Mechanism of depolymerization and severing of actin filaments and its significance in cytoskeletal dynamics. , 2007, International review of cytology.

[33]  A. Shevchenko,et al.  Direct Involvement of Yeast Type I Myosins in Cdc42-Dependent Actin Polymerization , 2000, The Journal of cell biology.

[34]  J. Gelles,et al.  Three-color single molecule imaging shows WASP detachment from Arp2/3 complex triggers actin filament branch formation , 2013, eLife.

[35]  A. Makhov,et al.  Coronin 1B Antagonizes Cortactin and Remodels Arp2/3-Containing Actin Branches in Lamellipodia , 2008, Cell.

[36]  H. Riezman,et al.  A novel EH domain protein of Saccharomyces cerevisiae, Ede1p, involved in endocytosis. , 2000, Journal of cell science.

[37]  Corbin E. Meacham,et al.  In Vivo RNAi Screening Identifies Regulators of Actin Dynamics as Key Determinants of Lymphoma Progression , 2009, Nature Genetics.

[38]  J. Condeelis,et al.  Bundling of actin filaments by elongation factor 1 alpha inhibits polymerization at filament ends , 1996, The Journal of cell biology.

[39]  J. Cooper,et al.  The role of Saccharomyces cerevisiae coronin in the actin and microtubule cytoskeletons , 1998, Current Biology.

[40]  S. Emr,et al.  Arrestin-Related Ubiquitin-Ligase Adaptors Regulate Endocytosis and Protein Turnover at the Cell Surface , 2008, Cell.

[41]  D. King,et al.  Arp2/3 complex is bound and activated by two WASP proteins , 2011, Proceedings of the National Academy of Sciences.

[42]  Daniel J. Lew,et al.  Inhibitory GEF Phosphorylation Provides Negative Feedback in the Yeast Polarity Circuit , 2014, Current Biology.

[43]  T. Tan,et al.  A Novel Src Homology 3 Domain-containing Adaptor Protein, HIP-55, That Interacts with Hematopoietic Progenitor Kinase 1* , 1999, The Journal of Biological Chemistry.

[44]  Niels Volkmann,et al.  The structural basis of actin filament branching by the Arp2/3 complex , 2008, The Journal of cell biology.

[45]  J. A. Gorman,et al.  Regulation of Profilin Localization in Saccharomyces cerevisiae by Phosphoinositide Metabolism (*) , 1995, The Journal of Biological Chemistry.

[46]  D. Drubin,et al.  Yeast Eps15-like endocytic protein, Pan1p, activates the Arp2/3 complex , 2001, Nature Cell Biology.

[47]  K. Ayscough,et al.  Under Pressure: the Differential Requirements for Actin during Yeast and Mammalian Endocytosis , 2009, Nature Cell Biology.

[48]  J. Keen,et al.  Interaction of phosphoinositide cycle intermediates with the plasma membrane-associated clathrin assembly protein AP-2. , 1991, The Journal of biological chemistry.

[49]  Bo Huang,et al.  Scd5p mediates phosphoregulation of actin and endocytosis by the type 1 phosphatase Glc7p in yeast. , 2007, Molecular biology of the cell.

[50]  J. Cooper,et al.  Disruption of the actin cytoskeleton in yeast capping protein mutants , 1990, Nature.

[51]  Justin M Keener,et al.  Quality Control and Substrate‐Dependent Downregulation of the Nutrient Transporter Fur4 , 2013, Traffic.

[52]  B. Haarer,et al.  Coordination of the filament stabilizing versus destabilizing activities of cofilin through its secondary binding site on actin , 2014, Cytoskeleton.

[53]  B. Goode,et al.  Functional Surfaces on the p35/ARPC2 Subunit of Arp2/3 Complex Required for Cell Growth, Actin Nucleation, and Endocytosis* , 2008, Journal of Biological Chemistry.

[54]  C. Volland,et al.  Endocytose and degradation of the uracil permease ofS. cerevisiae under stress conditions: Possible role of ubiquitin , 2008, Folia Microbiologica.

[55]  T. Pollard,et al.  Control of the Assembly of ATP- and ADP-Actin by Formins and Profilin , 2006, Cell.

[56]  C. Kaiser,et al.  Amino acids regulate retrieval of the yeast general amino acid permease from the vacuolar targeting pathway. , 2006, Molecular biology of the cell.

[57]  L. Blanchoin,et al.  Cofilin Tunes the Nucleotide State of Actin Filaments and Severs at Bare and Decorated Segment Boundaries , 2011, Current Biology.

[58]  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.

[59]  Michael N. Hall,et al.  Cell Wall Stress Depolarizes Cell Growth via Hyperactivation of Rho1 , 1999, The Journal of cell biology.

[60]  M. Whiteway,et al.  Forward genetics in Candida albicans that reveals the Arp2/3 complex is required for hyphal formation, but not endocytosis , 2010, Molecular microbiology.

[61]  Thomas D. Pollard,et al.  Structural and biochemical characterization of two binding sites for nucleation-promoting factor WASp-VCA on Arp2/3 complex , 2011, Proceedings of the National Academy of Sciences.

[62]  M. Hall,et al.  Sphingoid base signaling via Pkh kinases is required for endocytosis in yeast , 2001, The EMBO journal.

[63]  B. Wendland,et al.  Endocytic adaptors – social networking at the plasma membrane , 2011, Journal of Cell Science.

[64]  Thomas D. Pollard,et al.  Actin, a Central Player in Cell Shape and Movement , 2009, Science.

[65]  S. Winder,et al.  SCP1 encodes an actin-bundling protein in yeast. , 2003, The Biochemical journal.

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

[67]  T. Pollard,et al.  Interactions of WASp, myosin-I, and verprolin with Arp2/3 complex during actin patch assembly in fission yeast , 2005, The Journal of cell biology.

[68]  Zhaohui S. Qin,et al.  A Global Protein Kinase and Phosphatase Interaction Network in Yeast , 2010, Science.

[69]  Iwona I. Smaczynska-de Rooij,et al.  Interactions between the Yeast SM22 Homologue Scp1 and Actin Demonstrate the Importance of Actin Bundling in Endocytosis , 2008, Journal of Biological Chemistry.

[70]  T. Pollard,et al.  Synergies between Aip1p and capping protein subunits (Acp1p and Acp2p) in clathrin-mediated endocytosis and cell polarization in fission yeast , 2014, Molecular biology of the cell.

[71]  Ivan Dikic,et al.  CIN85 Participates in Cbl-b-mediated Down-regulation of Receptor Tyrosine Kinases* , 2002, The Journal of Biological Chemistry.

[72]  T. Pollard,et al.  Profilin binding to poly-L-proline and actin monomers along with ability to catalyze actin nucleotide exchange is required for viability of fission yeast. , 2001, Molecular biology of the cell.

[73]  Gary D. Bader,et al.  Bayesian Modeling of the Yeast SH3 Domain Interactome Predicts Spatiotemporal Dynamics of Endocytosis Proteins , 2009, PLoS biology.

[74]  T. Katoh,et al.  Actin-binding protein ABP140 is a methyltransferase for 3-methylcytidine at position 32 of tRNAs in Saccharomyces cerevisiae. , 2011, RNA.

[75]  D. Drubin,et al.  A Role for the Actin Cytoskeleton of Saccharomyces cerevisiae in Bipolar Bud-Site Selection , 1997, The Journal of cell biology.

[76]  Gianni Cesareni,et al.  Unusual Binding Properties of the SH3 Domain of the Yeast Actin-binding Protein Abp1 , 2002, The Journal of Biological Chemistry.

[77]  Bruce L. Goode,et al.  The Yeast Actin Cytoskeleton: from Cellular Function to Biochemical Mechanism , 2006, Microbiology and Molecular Biology Reviews.

[78]  Bo Huang,et al.  Identification of novel recognition motifs and regulatory targets for the yeast actin-regulating kinase Prk1p. , 2003, Molecular biology of the cell.

[79]  M. Balasubramanian,et al.  The novel fission yeast protein Pal1p interacts with Hip1-related Sla2p/End4p and is involved in cellular morphogenesis. , 2005, Molecular biology of the cell.

[80]  R. T. Hudson,et al.  Scd5p and clathrin function are important for cortical actin organization, endocytosis, and localization of sla2p in yeast. , 2002, Molecular biology of the cell.

[81]  C. Brautigam,et al.  Hierarchical regulation of WASP/WAVE proteins. , 2008, Molecular cell.

[82]  M. Geli,et al.  Crosstalk between PI(4,5)P₂and CK2 modulates actin polymerization during endocytic uptake. , 2014, Developmental cell.

[83]  Sandra L Schmid,et al.  Cargo and Dynamin Regulate Clathrin-Coated Pit Maturation , 2009, PLoS biology.

[84]  Howard Riezman,et al.  The F-Box Protein Rcy1p Is Involved in Endocytic Membrane Traffic and Recycling Out of an Early Endosome in Saccharomyces cerevisiae , 2000, The Journal of cell biology.

[85]  B. Winsor,et al.  Yeast Rsp5 ubiquitin ligase affects the actin cytoskeleton in vivo and in vitro. , 2011, European journal of cell biology.

[86]  G. Vaduva,et al.  Actin-binding Verprolin Is a Polarity Development Protein Required for the Morphogenesis and Function of the Yeast Actin Cytoskeleton , 1997, The Journal of cell biology.

[87]  L. Pon,et al.  Actin cable dynamics in budding yeast , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[88]  A. Spang,et al.  Cotranslational transport of ABP140 mRNA to the distal pole of S. cerevisiae , 2011, The EMBO journal.

[89]  T. Graham,et al.  Role for Drs2p, a P-Type Atpase and Potential Aminophospholipid Translocase, in Yeast Late Golgi Function , 1999, The Journal of cell biology.

[90]  Michael R Dores,et al.  Interaction between Epsin/Yap180 adaptors and the scaffolds Ede1/Pan1 is required for endocytosis. , 2008, Molecular biology of the cell.

[91]  M. Aigle,et al.  Rvs161p and Sphingolipids Are Required for Actin Repolarization following Salt Stress , 2002, Eukaryotic Cell.

[92]  T. Pollard,et al.  Direct observation of dendritic actin filament networks nucleated by Arp2/3 complex and WASP/Scar proteins , 2000, Nature.

[93]  D. Drubin,et al.  Clathrin-mediated endocytosis in budding yeast. , 2012, Trends in cell biology.

[94]  Fred Chang,et al.  Characterization of Dip1p Reveals a Switch in Arp2/3-Dependent Actin Assembly for Fission Yeast Endocytosis , 2011, Current Biology.

[95]  James Moseley,et al.  An actin nucleation mechanism mediated by Bni1 and Profilin , 2002, Nature Cell Biology.

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

[97]  T. Pollard,et al.  Interaction of Actin Monomers with AcanthamoebaActophorin (ADF/Cofilin) and Profilin* , 1998, The Journal of Biological Chemistry.

[98]  T. Pollard,et al.  The Role of the FH1 Domain and Profilin in Formin-Mediated Actin-Filament Elongation and Nucleation , 2008, Current Biology.

[99]  Charles Boone,et al.  A Role for Myosin-I in Actin Assembly through Interactions with Vrp1p, Bee1p, and the Arp2/3 Complex , 2000, The Journal of cell biology.

[100]  B. Wendland,et al.  PtdIns(3,5)P2 is Required for Delivery of Endocytic Cargo into the Multivesicular Body , 2003, Traffic.

[101]  M. Foti,et al.  Distinct roles for the yeast phosphatidylinositol 4-kinases, Stt4p and Pik1p, in secretion, cell growth, and organelle membrane dynamics. , 2000, Molecular biology of the cell.

[102]  Max A. Horlbeck,et al.  ER Cargo Properties Specify a Requirement for COPII Coat Rigidity Mediated by Sec13p , 2012, Science.

[103]  Cristian Suarez,et al.  Homeostatic Actin Cytoskeleton Networks Are Regulated by Assembly Factor Competition for Monomers , 2014, Current Biology.

[104]  H. Riezman,et al.  Two yeast mutants defective in endocytosis are defective in pheromone response , 1986, Cell.

[105]  P. De Camilli,et al.  Synaptojanin family members are implicated in endocytic membrane traffic in yeast. , 1998, Journal of cell science.

[106]  Michael Costanzo,et al.  Actin filament elongation in Arp2/3-derived networks is controlled by three distinct mechanisms. , 2013, Developmental cell.

[107]  G. Raposo,et al.  A dual role for K63-linked ubiquitin chains in multivesicular body biogenesis and cargo sorting , 2012, Molecular biology of the cell.

[108]  David G. Drubin,et al.  Determinants of endocytic membrane geometry, stability, and scission , 2011, Proceedings of the National Academy of Sciences.

[109]  A. Motley,et al.  Clathrin-mediated endocytosis in AP-2–depleted cells , 2003, The Journal of cell biology.

[110]  W. Marshall,et al.  Analysis of Clathrin-mediated Endocytosis of Epidermal Growth Factor Receptor by RNA Interference*[boxs] , 2004, Journal of Biological Chemistry.

[111]  T. Graham Flippases and vesicle-mediated protein transport. , 2004, Trends in cell biology.

[112]  Thomas D. Pollard,et al.  Mathematical Modeling of Endocytic Actin Patch Kinetics in Fission Yeast: Disassembly Requires Release of Actin Filament Fragments , 2010, Molecular biology of the cell.

[113]  P. Lappalainen,et al.  Cofilin promotes rapid actin filament turnover in vivo , 1997, Nature.

[114]  D. Boettner,et al.  Lessons from yeast for clathrin-mediated endocytosis , 2011, Nature Cell Biology.

[115]  Dan Wang,et al.  Drosophila twinfilin is required for cell migration and synaptic endocytosis , 2010, Journal of Cell Science.

[116]  S. Desrivières,et al.  MSS4, a Phosphatidylinositol-4-phosphate 5-Kinase Required for Organization of the Actin Cytoskeleton in Saccharomyces cerevisiae * , 1998, The Journal of Biological Chemistry.

[117]  Daniel J. Lew,et al.  A Morphogenesis Checkpoint Monitors the Actin Cytoskeleton in Yeast , 1998, The Journal of cell biology.

[118]  L. Blanchoin,et al.  Autonomous and in trans functions for the two halves of Srv2/CAP in promoting actin turnover , 2014, Cytoskeleton.

[119]  J. Gerst,et al.  Yeast exocytic v-SNAREs confer endocytosis. , 2000, Molecular biology of the cell.

[120]  T. Obinata,et al.  XAIP1: a Xenopus homologue of yeast actin interacting protein 1 (AIP1), which induces disassembly of actin filaments cooperatively with ADF/cofilin family proteins. , 1999, Journal of cell science.

[121]  B. Goode,et al.  Actin nucleation and elongation factors: mechanisms and interplay. , 2009, Current opinion in cell biology.

[122]  B. Wendland Faculty Opinions recommendation of Novel cargo-binding site in the beta and delta subunits of coatomer. , 2007 .

[123]  E. D. de Hostos The coronin family of actin-associated proteins. , 1999, Trends in cell biology.

[124]  M. Aigle,et al.  Cloning of the Multicopy Suppressor Gene SUR7: Evidence for a Functional Relationship between the Yeast Actin‐binding Protein Rvs167 and a Putative Membranous Protein , 1998, Yeast.

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

[126]  A. Goldman,et al.  Structure of the actin-depolymerizing factor homology domain in complex with actin , 2008, The Journal of cell biology.

[127]  Masako Osumi,et al.  Three-dimensional arrangement of F-actin in the contractile ring of fission yeast , 2007, The Journal of cell biology.

[128]  S. Emr,et al.  The yeast synaptojanin-like proteins control the cellular distribution of phosphatidylinositol (4,5)-bisphosphate. , 2002, Molecular biology of the cell.

[129]  K. Shianna,et al.  Tissue-Specific Genetic Control of Splicing: Implications for the Study of Complex Traits , 2008, PLoS biology.

[130]  B. Wendland,et al.  The Yeast Epsin Ent1 Is Recruited to Membranes through Multiple Independent Interactions* , 2003, The Journal of Biological Chemistry.

[131]  Daniel Feliciano,et al.  SLAC, a complex between Sla1 and Las17, regulates actin polymerization during clathrin-mediated endocytosis , 2012, Molecular biology of the cell.

[132]  A. Rodal,et al.  Structural and functional dissection of the Abp 1 ADFH actin-binding domain reveals versatile in vivo adapter functions , 2005 .

[133]  C. Schutt,et al.  The structure of crystalline profilin-beta-actin. , 1993, Nature.

[134]  H. Pelham,et al.  Slow Diffusion of Proteins in the Yeast Plasma Membrane Allows Polarity to Be Maintained by Endocytic Cycling , 2003, Current Biology.

[135]  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.

[136]  Malte Wachsmuth,et al.  Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis , 2012, Proceedings of the National Academy of Sciences.

[137]  U. Lindberg,et al.  Specific interaction between phosphatidylinositol 4,5-bisphosphate and profilactin , 1985, Nature.

[138]  Adam C. Martin,et al.  Critical conformational changes in the Arp2/3 complex are induced by nucleotide and nucleation promoting factor. , 2004, Molecular cell.

[139]  P. Evans,et al.  A structural explanation for the recognition of tyrosine-based endocytotic signals. , 1998, Science.

[140]  H. Riezman,et al.  End4p/Sla2p interacts with actin-associated proteins for endocytosis in Saccharomyces cerevisiae. , 1997, Molecular biology of the cell.

[141]  T. Graham,et al.  Yeast P4-ATPases Drs2p and Dnf1p are essential cargos of the NPFXD/Sla1p endocytic pathway. , 2006, Molecular biology of the cell.

[142]  J. Mattila,et al.  GMF Promotes Leading-Edge Dynamics and Collective Cell Migration In Vivo , 2014, Current Biology.

[143]  J. Cooper,et al.  Roles for actin assembly in endocytosis. , 2012, Annual review of biochemistry.

[144]  A. Munn,et al.  EH domain proteins Pan1p and End3p are components of a complex that plays a dual role in organization of the cortical actin cytoskeleton and endocytosis in Saccharomyces cerevisiae , 1997, Molecular and cellular biology.

[145]  J. Hesketh Translation and the cytoskeleton: a mechanism for targeted protein synthesis , 1994, Molecular Biology Reports.

[146]  Hsin-Yao Tang,et al.  Pan1p, End3p, and Sla1p, Three Yeast Proteins Required for Normal Cortical Actin Cytoskeleton Organization, Associate with Each Other and Play Essential Roles in Cell Wall Morphogenesis , 2000, Molecular and Cellular Biology.

[147]  P. Camilli,et al.  A presynaptic inositol-5-phosphatase , 1996, Nature.

[148]  M. Wigler,et al.  Cloning and characterization of CAP, the S. cerevisiae gene encoding the 70 kd adenylyl cyclase-associated protein , 1990, Cell.

[149]  F. Chang,et al.  Role of turgor pressure in endocytosis in fission yeast , 2014, Molecular biology of the cell.

[150]  D. Sept,et al.  A genetic dissection of Aip1p's interactions leads to a model for Aip1p-cofilin cooperative activities. , 2006, Molecular biology of the cell.

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

[152]  K. Ayscough,et al.  A Novel Actin-Binding Motif in Las17/WASP Nucleates Actin Filaments Independently of Arp2/3 , 2013, Current Biology.

[153]  J. Bartles Parallel actin bundles and their multiple actin-bundling proteins. , 2000, Current opinion in cell biology.

[154]  Adam C. Martin,et al.  Arp2/3 ATP hydrolysis-catalysed branch dissociation is critical for endocytic force generation , 2006, Nature Cell Biology.

[155]  Marie-France Carlier,et al.  Reconstitution of actin-based motility of Listeria and Shigella using pure proteins , 1999, Nature.

[156]  M. Gimona,et al.  Calponins: adaptable modular regulators of the actin cytoskeleton. , 2008, The international journal of biochemistry & cell biology.

[157]  G. Payne,et al.  Adaptor complex-independent clathrin function in yeast. , 1999, Molecular biology of the cell.

[158]  S. Schmid,et al.  Identification of an adaptor-associated kinase, AAK1, as a regulator of clathrin-mediated endocytosis , 2002, The Journal of cell biology.

[159]  F. Chang Movement of a cytokinesis factor cdc12p to the site of cell division , 1999, Current Biology.

[160]  Niels Volkmann,et al.  A protein disulfide oxidoreductase from the archaeon Pyrococcus furiosus contains two thioredoxin fold units , 1998, Nature Structural Biology.

[161]  T. Kinzy,et al.  Translation elongation factor 1A is essential for regulation of the actin cytoskeleton and cell morphology , 2005, Nature Structural &Molecular Biology.

[162]  Niels Volkmann,et al.  An Atomic Model of Actin Filaments Cross-Linked by Fimbrin and Its Implications for Bundle Assembly and Function , 2001, The Journal of cell biology.

[163]  B. André,et al.  Internal Amino Acids Promote Gap1 Permease Ubiquitylation via TORC1/Npr1/14-3-3-Dependent Control of the Bul Arrestin-Like Adaptors , 2012, Molecular and Cellular Biology.

[164]  Clathrin Coat Disassembly by the Yeast Hsc70/Ssa1p and Auxilin/Swa2p Proteins Observed by Single-particle Burst Analysis Spectroscopy* , 2013, The Journal of Biological Chemistry.

[165]  W. Brieher,et al.  Cyclase-associated Protein (CAP) Acts Directly on F-actin to Accelerate Cofilin-mediated Actin Severing across the Range of Physiological pH* , 2012, The Journal of Biological Chemistry.

[166]  Rong Li,et al.  Activation of the yeast Arp2/3 complex by Bee1p, a WASP-family protein , 1999, Current Biology.

[167]  J. Cooper,et al.  Distinct Roles for Arp2/3 Regulators in Actin Assembly and Endocytosis , 2008, PLoS biology.

[168]  D. Drubin,et al.  Regulation of cortical actin cytoskeleton assembly during polarized cell growth in budding yeast , 1995, The Journal of cell biology.

[169]  J. Gelles,et al.  Pathway of actin filament branch formation by Arp2/3 complex revealed by single-molecule imaging , 2013, Proceedings of the National Academy of Sciences.

[170]  K. Ayscough,et al.  An Abp1-Dependent Route of Endocytosis Functions when the Classical Endocytic Pathway in Yeast Is Inhibited , 2014, PloS one.

[171]  H. Kueh,et al.  Actin disassembly by cofilin, coronin, and Aip1 occurs in bursts and is inhibited by barbed-end cappers , 2008, The Journal of cell biology.

[172]  G. Payne,et al.  Structure of Sla1p homology domain 1 and interaction with the NPFxD endocytic internalization motif , 2007, The EMBO journal.

[173]  D. Drubin,et al.  Evidence for physical and functional interactions among two Saccharomyces cerevisiae SH3 domain proteins, an adenylyl cyclase-associated protein and the actin cytoskeleton. , 1997, Molecular biology of the cell.

[174]  Francesca Santini,et al.  Spatial control of coated-pit dynamics in living cells , 1999, Nature Cell Biology.

[175]  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.

[176]  H. Riezman,et al.  Identification and characterization of Saccharomyces cerevisiae mutants defective in fluid‐phase endocytosis , 2001, Yeast.

[177]  A. Nakano,et al.  Cdc 50 p , a Conserved Endosomal Membrane Protein , Controls Polarized Growth in Saccharomyces cerevisiae , 2003 .

[178]  Kartik Chandran,et al.  Endocytosis by Random Initiation and Stabilization of Clathrin-Coated Pits , 2004, Cell.

[179]  Omar Quintero-Monzon,et al.  A central role for the WH2 domain of Srv2/CAP in recharging actin monomers to drive actin turnover in vitro and in vivo , 2010, Cytoskeleton.

[180]  Iwona I. Smaczynska-de Rooij,et al.  A role for the dynamin-like protein Vps1 during endocytosis in yeast , 2010, Journal of Cell Science.

[181]  Philip V. Bayly,et al.  Force generation by endocytic actin patches in budding yeast. , 2014, Biophysical journal.

[182]  S. Gygi,et al.  Purification and architecture of the ubiquitous Wave complex. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[183]  A. Annila,et al.  Structural basis and evolutionary origin of actin filament capping by twinfilin , 2007, Proceedings of the National Academy of Sciences.

[184]  Matthew D. Welch,et al.  A nucleator arms race: cellular control of actin assembly , 2010, Nature Reviews Molecular Cell Biology.

[185]  Gianni Cesareni,et al.  Recognition specificity of individual EH domains of mammals and yeast , 1998, The EMBO journal.

[186]  P. P. Di Fiore,et al.  Nucleocytoplasmic Shuttling of Endocytic Proteins , 2001, The Journal of cell biology.

[187]  H. Riezman,et al.  Characterization of the END1 gene required for vacuole biogenesis and gluconeogenic growth of budding yeast. , 1989, The EMBO journal.

[188]  G. Payne,et al.  The sequence NPFXD defines a new class of endocytosis signal in Saccharomyces cerevisiae , 1996, The Journal of cell biology.

[189]  D. Portnoy,et al.  Actin filaments and the growth, movement, and spread of the intracellular bacterial parasite, Listeria monocytogenes , 1989, The Journal of cell biology.

[190]  M. Aigle,et al.  Actin cytoskeleton and budding pattern are altered in the yeast rvs161 mutant: the Rvs161 protein shares common domains with the brain protein amphiphysin , 1995, Molecular and General Genetics MGG.

[191]  K. Ayscough,et al.  Nucleocytoplasmic Trafficking Is Required for Functioning of the Adaptor Protein Sla1p in Endocytosis , 2007, Traffic.

[192]  E. L. Hostos,et al.  The coronin family of actin-associated proteins , 1999 .

[193]  Y. Takai,et al.  Interaction of Rho1p target Bni1p with F-actin-binding elongation factor 1α: implication in Rho1p-regulated reorganization of the actin cytoskeleton in Saccharomyces cerevisiae , 1998, Oncogene.

[194]  Kenneth R. Henry,et al.  Cortical recruitment and nuclear-cytoplasmic shuttling of Scd5p, a protein phosphatase-1-targeting protein involved in actin organization and endocytosis. , 2005, Molecular biology of the cell.

[195]  G. Schröder,et al.  Remodeling of actin filaments by ADF/cofilin proteins , 2011, Proceedings of the National Academy of Sciences.

[196]  Gary D Bader,et al.  The Genetic Landscape of a Cell , 2010, Science.

[197]  P. Bork,et al.  Functional organization of the yeast proteome by systematic analysis of protein complexes , 2002, Nature.

[198]  Petri Auvinen,et al.  Mouse A6/Twinfilin Is an Actin Monomer-Binding Protein That Localizes to the Regions of Rapid Actin Dynamics , 2000, Molecular and Cellular Biology.

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

[200]  E. Eisenberg,et al.  Role of Cyclin G-associated Kinase in Uncoating Clathrin-coated Vesicles from Non-neuronal Cells* , 2000, The Journal of Biological Chemistry.

[201]  Robert H. Insall,et al.  F-BAR domains: multifunctional regulators of membrane curvature , 2008, Journal of Cell Science.

[202]  D. Drubin,et al.  Cofilin recruitment and function during actin-mediated endocytosis dictated by actin nucleotide state , 2007, The Journal of cell biology.

[203]  Howard Riezman,et al.  Distinct acto/myosin-I structures associate with endocytic profiles at the plasma membrane , 2008, The Journal of cell biology.

[204]  C. Landry,et al.  An in Vivo Map of the Yeast Protein Interactome , 2008, Science.

[205]  John A.G. Briggs,et al.  Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision , 2011, The Journal of cell biology.

[206]  E. W. Jones,et al.  Clathrin requirement for normal growth of yeast. , 1987, Science.

[207]  T. Stevens,et al.  The VPS1 protein, a homolog of dynamin required for vacuolar protein sorting in Saccharomyces cerevisiae, is a GTPase with two functionally separable domains , 1992, The Journal of cell biology.

[208]  R. Kölling,et al.  The ABC‐transporter Ste6 accumulates in the plasma membrane in a ubiquitinated form in endocytosis mutants. , 1994, The EMBO journal.

[209]  T. Pollard,et al.  Influence of the C terminus of Wiskott-Aldrich syndrome protein (WASp) and the Arp2/3 complex on actin polymerization. , 1999, Biochemistry.

[210]  Rong Li,et al.  Bee1, a Yeast Protein with Homology to Wiscott-Aldrich Syndrome Protein, Is Critical for the Assembly of Cortical Actin Cytoskeleton , 1997, The Journal of cell biology.

[211]  B. Goode,et al.  The p40/ARPC1 Subunit of Arp2/3 Complex Performs Multiple Essential Roles in WASp-regulated Actin Nucleation* , 2010, The Journal of Biological Chemistry.

[212]  S. Schmid,et al.  Endocytosis: Past, present, and future. , 2014, Cold Spring Harbor perspectives in biology.

[213]  P. Rubenstein,et al.  An Intermediate Form of ADP-F-actin* , 2005, Journal of Biological Chemistry.

[214]  A. DePace,et al.  The Human Arp2/3 Complex Is Composed of Evolutionarily Conserved Subunits and Is Localized to Cellular Regions of Dynamic Actin Filament Assembly , 1997, The Journal of cell biology.

[215]  O. Sokolova,et al.  GMF Severs Actin-Arp2/3 Complex Branch Junctions by a Cofilin-like Mechanism , 2013, Current Biology.

[216]  R. Knorr,et al.  Unusual Structural Organization of the Endocytic Proteins AP180 and Epsin 1* , 2002, The Journal of Biological Chemistry.

[217]  B. Haarer,et al.  Structure determination and characterization of Saccharomyces cerevisiae profilin. , 1998, Biochemistry.

[218]  B. Kay,et al.  Characterizing Class I WW domains defines key specificity determinants and generates mutant domains with novel specificities. , 2001, Chemistry & biology.

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

[220]  W. Frommer,et al.  Plasma membrane microdomains regulate turnover of transport proteins in yeast , 2008, The Journal of cell biology.

[221]  D. Drubin,et al.  Analysis of yeast endocytic site formation and maturation through a regulatory transition point , 2012, Molecular biology of the cell.

[222]  S. Emr,et al.  Yeast epsins contain an essential N‐terminal ENTH domain, bind clathrin and are required for endocytosis , 1999, The EMBO journal.

[223]  T D Pollard,et al.  Rate constants for the reactions of ATP- and ADP-actin with the ends of actin filaments , 1986, The Journal of cell biology.

[224]  T. Pollard,et al.  Distinct Roles for F-BAR Proteins Cdc15p and Bzz1p in Actin Polymerization at Sites of Endocytosis in Fission Yeast , 2011, Current Biology.

[225]  D. Sept,et al.  An actin-filament-binding interface on the Arp2/3 complex is critical for nucleation and branch stability , 2010, Proceedings of the National Academy of Sciences.

[226]  Felix Campelo,et al.  Membrane fission: the biogenesis of transport carriers. , 2012, Annual review of biochemistry.

[227]  Kyoungtae Kim,et al.  Inactivation of Tor proteins affects the dynamics of endocytic proteins in early stage of endocytosis , 2013, Journal of Biosciences.

[228]  T. Pollard,et al.  Scar, a WASp-related protein, activates nucleation of actin filaments by the Arp2/3 complex. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[229]  P. Lappalainen,et al.  Actin‐depolymerizing factor homology domain: A conserved fold performing diverse roles in cytoskeletal dynamics , 2011, Cytoskeleton.

[230]  S. Kohlwein,et al.  Phospholipid synthesis and lipid composition of subcellular membranes in the unicellular eukaryote Saccharomyces cerevisiae , 1991, Journal of bacteriology.

[231]  O. Sokolova,et al.  Structure and Mechanism of Mouse Cyclase-associated Protein (CAP1) in Regulating Actin Dynamics* , 2014, The Journal of Biological Chemistry.

[232]  A. Dautry‐Varsat,et al.  Enhancement of endocytosis due to aminophospholipid transport across the plasma membrane of living cells. , 1999, The American journal of physiology.

[233]  S. Emr,et al.  TORC1 Regulates Endocytosis via Npr1-Mediated Phosphoinhibition of a Ubiquitin Ligase Adaptor , 2011, Cell.

[234]  T. Pollard,et al.  Twinfilin is an actin-filament-severing protein and promotes rapid turnover of actin structures in vivo , 2006, Journal of Cell Science.

[235]  Thomas M. Newpher,et al.  In vivo dynamics of clathrin and its adaptor-dependent recruitment to the actin-based endocytic machinery in yeast. , 2005, Developmental cell.

[236]  D. Drubin,et al.  Multiple Pathways Regulate Endocytic Coat Disassembly in Saccharomyces cerevisiae for Optimal Downstream Trafficking , 2008, Traffic.

[237]  David Botstein,et al.  Defining protein interactions with yeast actin in vivo , 1995, Nature Structural Biology.

[238]  B. Andrews,et al.  Direct regulation of Arp2/3 complex activity and function by the actin binding protein coronin , 2002, The Journal of cell biology.

[239]  P. Janmey,et al.  Cofilin is an essential component of the yeast cortical cytoskeleton , 1993, The Journal of cell biology.

[240]  R. Schneiter,et al.  The Sur7p Family Defines Novel Cortical Domains in Saccharomyces cerevisiae, Affects Sphingolipid Metabolism, and Is Involved in Sporulation , 2002, Molecular and Cellular Biology.

[241]  S. Almo,et al.  A high-affinity interaction with ADP-actin monomers underlies the mechanism and in vivo function of Srv2/cyclase-associated protein. , 2004, Molecular biology of the cell.

[242]  B. Nolen,et al.  Structural basis for regulation of Arp2/3 complex by GMF , 2013, Nature Structural &Molecular Biology.

[243]  David G. Drubin,et al.  Novel Protein Kinases Ark1p and Prk1p Associate with and Regulate the Cortical Actin Cytoskeleton in Budding Yeast , 1999, The Journal of cell biology.

[244]  Thomas M. Newpher,et al.  Clathrin is Important for Normal Actin Dynamics and Progression of Sla2p‐Containing Patches During Endocytosis in Yeast , 2006, Traffic.

[245]  T. Maritzen,et al.  Turning CALM into excitement: AP180 and CALM in endocytosis and disease , 2012, Biology of the cell.

[246]  B. Wendland,et al.  Targeted Disruption of an EH‐domain Protein Endocytic Complex, Pan1‐End3 , 2014, Traffic.

[247]  D. Baker,et al.  Protein transport to the vacuole and receptor-mediated endocytosis by clathrin heavy chain-deficient yeast , 1988, The Journal of cell biology.

[248]  E. O’Shea,et al.  Global analysis of protein localization in budding yeast , 2003, Nature.

[249]  D R Bentley,et al.  Genomic Sequence Information Should Be Released Immediately and Freely in the Public Domain , 1996, Science.

[250]  Scott D. Emr,et al.  Pan1p, Yeast eps15, Functions as a Multivalent Adaptor That Coordinates Protein–Protein Interactions Essential for Endocytosis , 1998, The Journal of cell biology.

[251]  T. Pollard,et al.  Mechanism of Interaction of Acanthamoeba Actophorin (ADF/Cofilin) with Actin Filaments* , 1999, The Journal of Biological Chemistry.

[252]  F. Aguet,et al.  The First Five Seconds in the Life of a Clathrin-Coated Pit , 2012, Cell.

[253]  J. Cooper,et al.  Actin-based motility during endocytosis in budding yeast. , 2005, Molecular biology of the cell.

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

[255]  T. Pollard,et al.  Actin' like actin? , 1996, Trends in cell biology.

[256]  Verónica A. Segarra,et al.  Role of Scd5, a protein phosphatase-1 targeting protein, in phosphoregulation of Sla1 during endocytosis , 2012, Journal of Cell Science.

[257]  T. Pollard,et al.  Identification of functionally important residues of Arp2/3 complex by analysis of homology models from diverse species. , 2004, Journal of molecular biology.

[258]  Thomas D. Pollard,et al.  Actin Filament Severing by Cofilin Dismantles Actin Patches and Produces Mother Filaments for New Patches , 2013, Current Biology.

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

[260]  B. Pearse Coated vesicles from pig brain: purification and biochemical characterization. , 1975, Journal of molecular biology.

[261]  A. Rodal,et al.  Actin and septin ultrastructures at the budding yeast cell cortex. , 2004, Molecular Biology of the Cell.

[262]  T. Pollard,et al.  Purification of a cortical complex containing two unconventional actins from Acanthamoeba by affinity chromatography on profilin-agarose , 1994, The Journal of cell biology.

[263]  D. Sept,et al.  New insights into mechanism and regulation of actin capping protein. , 2008, International review of cell and molecular biology.

[264]  Anita T. Layton,et al.  Modeling Vesicle Traffic Reveals Unexpected Consequences for Cdc42p-Mediated Polarity Establishment , 2011, Current Biology.

[265]  E. Freire,et al.  Epsin N-terminal homology domains perform an essential function regulating Cdc42 through binding Cdc42 GTPase-activating proteins. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[266]  D. Shaywitz,et al.  2 Protein Secretion, Membrane Biogenesis, and Endocytosis , 1997 .

[267]  M. Kaksonen,et al.  Harnessing actin dynamics for clathrin-mediated endocytosis , 2006, Nature Reviews Molecular Cell Biology.

[268]  D. Drubin,et al.  Early-arriving Syp1p and Ede1p function in endocytic site placement and formation in budding yeast. , 2009, Molecular biology of the cell.

[269]  Samuel H. Payne,et al.  A Multidimensional Chromatography Technology for In-depth Phosphoproteome Analysis*S , 2008, Molecular & Cellular Proteomics.

[270]  Laurent Blanchoin,et al.  Coronin switches roles in actin disassembly depending on the nucleotide state of actin. , 2009, Molecular cell.

[271]  Joshua D. Schnell,et al.  The Function of Yeast Epsin and Ede1 Ubiquitin‐Binding Domains During Receptor Internalization , 2010, Traffic.

[272]  Simon Hippenmeyer,et al.  Conformational changes in the Arp2/3 complex leading to actin nucleation , 2005, Nature Structural &Molecular Biology.

[273]  B. Winsor,et al.  The WASP/Las17p-interacting protein Bzz1p functions with Myo5p in an early stage of endocytosis , 2005, Protoplasma.

[274]  Thomas D. Pollard,et al.  Quantitative Analysis of the Mechanism of Endocytic Actin Patch Assembly and Disassembly in Fission Yeast , 2010, Molecular biology of the cell.

[275]  R. Schekman,et al.  Genetic and biochemical characterization of clathrin-deficient Saccharomyces cerevisiae , 1987, Molecular and cellular biology.

[276]  David G. Drubin,et al.  A Modular Design for the Clathrin- and Actin-Mediated Endocytosis Machinery , 2005, Cell.

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

[278]  B. Wendland,et al.  The F-BAR Protein Syp1 Negatively Regulates WASp-Arp2/3 Complex Activity during Endocytic Patch Formation , 2009, Current Biology.

[279]  B. André,et al.  NPI1, an essential yeast gene involved in induced degradation of Gap1 and Fur4 permeases, encodes the Rsp5 ubiquitin—protein ligase , 1995, Molecular microbiology.

[280]  E. Phizicky,et al.  A domain of the actin binding protein Abp140 is the yeast methyltransferase responsible for 3-methylcytidine modification in the tRNA anti-codon loop. , 2011, RNA.

[281]  S. Winder,et al.  The WASP homologue Las17 activates the novel actin-regulatory activity of Ysc84 to promote endocytosis in yeast. , 2009, Molecular biology of the cell.

[282]  S. Keeney,et al.  References and Notes Supporting Online Material Materials and Methods Figs. S1 to S5 Tables S1 and S2 References Movie S1 Fcho Proteins Are Nucleators of Clathrin-mediated Endocytosis , 2022 .

[283]  B. Wendland,et al.  The Sla2p talin domain plays a role in endocytosis in Saccharomyces cerevisiae. , 2003, Genetics.

[284]  J. Condeelis,et al.  How is actin polymerization nucleated in vivo? , 2001, Trends in cell biology.

[285]  David G. Drubin,et al.  Building Distinct Actin Filament Networks in a Common Cytoplasm , 2011, Current Biology.

[286]  A. Shevchenko,et al.  Saccharomyces cerevisiae Bzz1p Is Implicated with Type I Myosins in Actin Patch Polarization and Is Able To Recruit Actin-Polymerizing Machinery In Vitro , 2002, Molecular and Cellular Biology.

[287]  Alexander Varshavsky,et al.  The ubiquitin system, an immense realm. , 2012, Annual review of biochemistry.

[288]  Walter E. Gall,et al.  The auxilin-like phosphoprotein Swa2p is required for clathrin function in yeast , 2000, Current Biology.

[289]  M. Kaksonen,et al.  Reassessment of the role of plasma membrane domains in the regulation of vesicular traffic in yeast , 2011, Journal of Cell Science.

[290]  J. Berman,et al.  SLA2 mutations cause SWE1-mediated cell cycle phenotypes in Candida albicans and Saccharomyces cerevisiae. , 2009, Microbiology.

[291]  T. Drivas,et al.  Existence of a novel clathrin-independent endocytic pathway in yeast that depends on Rho1 and formin , 2011, The Journal of cell biology.

[292]  Timothy J. Mitchison,et al.  Actin polymerization is induced by Arp 2/3 protein complex at the surface of Listeria monocytogenes , 1997, Nature.

[293]  L. Falquet,et al.  A ubiquitin-interacting motif conserved in components of the proteasomal and lysosomal protein degradation systems. , 2001, Trends in biochemical sciences.

[294]  H. Pelham,et al.  Inefficient Quality Control of Thermosensitive Proteins on the Plasma Membrane , 2009, PloS one.

[295]  Roberto Dominguez,et al.  Actin structure and function. , 2011, Annual review of biophysics.

[296]  S. Emr,et al.  A new vital stain for visualizing vacuolar membrane dynamics and endocytosis in yeast , 1995, The Journal of cell biology.

[297]  S. Confalonieri,et al.  EH and UIM: Endocytosis and More , 2003, Science's STKE.

[298]  P. Bork,et al.  A systematic screen for protein–lipid interactions in Saccharomyces cerevisiae , 2010, Molecular systems biology.

[299]  A. Sachs,et al.  Translation initiation requires the PAB-dependent poly(A) ribonuclease in yeast , 1992, Cell.

[300]  Anna Espinal,et al.  Ultrastructural dynamics of proteins involved in endocytic budding , 2012, Proceedings of the National Academy of Sciences.

[301]  H. Kuwayama,et al.  GMF is an evolutionarily developed Adf/cofilin‐super family protein involved in the Arp2/3 complex‐mediated organization of the actin cytoskeleton , 2010, Cytoskeleton.

[302]  B. Nolen,et al.  Dip1 Defines a Class of Arp2/3 Complex Activators that Function without Preformed Actin Filaments , 2013, Current Biology.

[303]  J. Cooper,et al.  Interactions with PIP2, ADP-actin monomers, and capping protein regulate the activity and localization of yeast twinfilin , 2001, The Journal of cell biology.

[304]  Walter E. Gall,et al.  Drs2p-Dependent Formation of Exocytic Clathrin-Coated Vesicles In Vivo , 2002, Current Biology.

[305]  H. Riezman,et al.  Clathrin functions in the absence of heterotetrameric adaptors and AP180‐related proteins in yeast , 1999, The EMBO journal.

[306]  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.

[307]  B. Peter,et al.  BAR Domains as Sensors of Membrane Curvature: The Amphiphysin BAR Structure , 2004, Science.

[308]  D. Kovar,et al.  Actin Filament Bundling by Fimbrin Is Important for Endocytosis, Cytokinesis, and Polarization in Fission Yeast* , 2011, The Journal of Biological Chemistry.

[309]  L. Hicke,et al.  A function for monoubiquitination in the internalization of a G protein-coupled receptor. , 1998, Molecular cell.

[310]  Yuan He,et al.  Ubiquitin binds to and regulates a subset of SH3 domains. , 2007, Molecular cell.

[311]  B. Wollscheid,et al.  SH3P7 Is a Cytoskeleton Adapter Protein and Is Coupled to Signal Transduction from Lymphocyte Antigen Receptors , 1999, Molecular and Cellular Biology.

[312]  N. Mueller,et al.  Patchwork organization of the yeast plasma membrane into numerous coexisting domains , 2012, Nature Cell Biology.

[313]  E. Jorgensen,et al.  Molecular basis of synaptic vesicle cargo recognition by the endocytic sorting adaptor stonin 2 , 2007, The Journal of cell biology.

[314]  G. Payne,et al.  NPFXD-mediated endocytosis is required for polarity and function of a yeast cell wall stress sensor. , 2006, Molecular biology of the cell.

[315]  A. Roth,et al.  Ubiquitination of the yeast a-factor receptor , 1996, The Journal of cell biology.

[316]  N. Barkai,et al.  The Competitive Advantage of a Dual-Transporter System , 2011, Science.

[317]  A. Nakano,et al.  Cdc50p, a conserved endosomal membrane protein, controls polarized growth in Saccharomyces cerevisiae. , 2003, Molecular biology of the cell.

[318]  T. Kinzy,et al.  Overexpression of translation elongation factor 1A affects the organization and function of the actin cytoskeleton in yeast. , 2001, Genetics.

[319]  James R. Broach,et al.  SRV2, a gene required for RAS activation of adenylate cyclase in yeast , 1990, Cell.

[320]  T. Pollard,et al.  Quantitative analysis of the effect of Acanthamoeba profilin on actin filament nucleation and elongation. , 1984, Biochemistry.

[321]  H. Riezman,et al.  Multiple functions of sterols in yeast endocytosis. , 2002, Molecular biology of the cell.

[322]  P. Mattila,et al.  Mechanism and biological role of profilin-Srv2/CAP interaction , 2007, Journal of Cell Science.

[323]  B. Haarer,et al.  Mutational analysis of yeast profilin , 1993, Molecular and cellular biology.

[324]  N. Nelson,et al.  Disruption of genes encoding subunits of yeast vacuolar H(+)-ATPase causes conditional lethality. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[325]  Anya L. Goodman,et al.  Coordinated Regulation of Actin Filament Turnover by a High-Molecular-Weight Srv2/CAP Complex, Cofilin, Profilin, and Aip1 , 2003, Current Biology.

[326]  Duilio Cascio,et al.  Regulation of clathrin adaptor function in endocytosis: novel role for the SAM domain , 2010, The EMBO journal.

[327]  M. Kaksonen,et al.  PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization , 2007, The Journal of cell biology.

[328]  M. Kirschner,et al.  The Interaction between N-WASP and the Arp2/3 Complex Links Cdc42-Dependent Signals to Actin Assembly , 1999, Cell.

[329]  T. Pollard,et al.  Xenopus Actin-interacting Protein 1 (XAip1) Enhances Cofilin Fragmentation of Filaments by Capping Filament Ends* , 2002, The Journal of Biological Chemistry.

[330]  Joshua D. Schnell,et al.  The conserved Pkh–Ypk kinase cascade is required for endocytosis in yeast , 2002, The Journal of cell biology.

[331]  P. Várnai,et al.  Live cell imaging of phosphoinositides with expressed inositide binding protein domains. , 2008, Methods.

[332]  H. Riezman,et al.  Specific sterols required for the internalization step of endocytosis in yeast. , 1999, Molecular biology of the cell.

[333]  B. Andrews,et al.  The Biologically Relevant Targets and Binding Affinity Requirements for the Function of the Yeast Actin-Binding Protein 1 Src-Homology 3 Domain Vary With Genetic Context , 2007, Genetics.

[334]  A. Fedorov,et al.  Essential functions and actin‐binding surfaces of yeast cofilin revealed by systematic mutagenesis , 1997, The EMBO journal.

[335]  T. Pollard,et al.  Mechanism of the interaction of human platelet profilin with actin , 1991, The Journal of cell biology.

[336]  G. Payne,et al.  A yeast DNA J protein required for uncoating of clathrin-coated vesicles in vivo , 2000, Nature Cell Biology.

[337]  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.

[338]  J. Yates,et al.  Reconstitution and Protein Composition Analysis of Endocytic Actin Patches , 2010, Current Biology.

[339]  Xianwen Yu,et al.  The yeast dynamin-related GTPase Vps1p functions in the organization of the actin cytoskeleton via interaction with Sla1p , 2004, Journal of Cell Science.

[340]  S. Lemmon,et al.  Suppressors of clathrin deficiency: overexpression of ubiquitin rescues lethal strains of clathrin-deficient Saccharomyces cerevisiae , 1993, Molecular and cellular biology.

[341]  D. Sept,et al.  Overlapping and distinct functions for cofilin, coronin and Aip1 in actin dynamics in vivo , 2010, Journal of Cell Science.

[342]  A. Fedorov,et al.  Identification of Functional Residues on Caenorhabditis elegans Actin-interacting Protein 1 (UNC-78) for Disassembly of Actin Depolymerizing Factor/Cofilin-bound Actin Filaments* , 2004, Journal of Biological Chemistry.

[343]  W. Almers,et al.  Imaging actin and dynamin recruitment during invagination of single clathrin-coated pits , 2002, Nature Cell Biology.

[344]  Pietro De Camilli,et al.  Dynamin, a membrane-remodelling GTPase , 2012, Nature Reviews Molecular Cell Biology.

[345]  B. Barrell,et al.  Life with 6000 Genes , 1996, Science.

[346]  D. Drubin,et al.  A yeast killer toxin screen provides insights into a/b toxin entry, trafficking, and killing mechanisms. , 2009, Developmental cell.

[347]  U. Liebel,et al.  Novel cargo-binding site in the β and δ subunits of coatomer , 2007, The Journal of cell biology.

[348]  B. Goode,et al.  Dissection of Arp2/3 Complex Actin Nucleation Mechanism and Distinct Roles for Its Nucleation-Promoting Factors in Saccharomyces cerevisiae , 2005, Genetics.

[349]  M. Rieger,et al.  The Yeast Pan2 Protein Is Required for Poly(A)-binding Protein-stimulated Poly(A)-nuclease Activity * , 1996, The Journal of Biological Chemistry.

[350]  Harvey T. McMahon,et al.  Molecular mechanism and physiological functions of clathrin-mediated endocytosis , 2011, Nature Reviews Molecular Cell Biology.

[351]  D. Kovar,et al.  Fimbrin and Tropomyosin Competition Regulates Endocytosis and Cytokinesis Kinetics in Fission Yeast , 2010, Current Biology.

[352]  A. Bretscher,et al.  Analysis of unregulated formin activity reveals how yeast can balance F-actin assembly between different microfilament-based organizations. , 2008, Molecular biology of the cell.

[353]  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.

[354]  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.

[355]  David G. Drubin,et al.  Activation of the Arp2/3 Complex by the Actin Filament Binding Protein Abp1p , 2001, The Journal of cell biology.

[356]  P. Mattila,et al.  Twinfilin is required for actin-dependent developmental processes in Drosophila , 2001, The Journal of cell biology.

[357]  S. Emr,et al.  The ART-Rsp5 ubiquitin ligase network comprises a plasma membrane quality control system that protects yeast cells from proteotoxic stress , 2013, eLife.

[358]  J. Jin,et al.  Calponin in Non-Muscle Cells , 2008, Cell Biochemistry and Biophysics.

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

[360]  D. Lew,et al.  Morphogenesis and the Cell Cycle , 2012, Genetics.

[361]  Joshua D. Schnell,et al.  Receptor internalization in yeast requires the Tor2-Rho1 signaling pathway. , 2003, Molecular biology of the cell.

[362]  Adam C. Martin,et al.  Endocytic internalization in budding yeast requires coordinated actin nucleation and myosin motor activity. , 2006, Developmental cell.

[363]  S. Emr,et al.  The dual PH domain protein Opy1 functions as a sensor and modulator of PtdIns(4,5)P2 synthesis , 2012, The EMBO journal.

[364]  D. Drubin,et al.  Interaction of the endocytic scaffold protein Pan1 with the type I myosins contributes to the late stages of endocytosis. , 2007, Molecular biology of the cell.

[365]  Harini Ravi,et al.  Aip1 and cofilin promote rapid turnover of yeast actin patches and cables: a coordinated mechanism for severing and capping filaments. , 2006, Molecular biology of the cell.

[366]  Mingjie Cai,et al.  Regulation of the Actin Cytoskeleton Organization in Yeast by a Novel Serine/Threonine Kinase Prk1p , 1999, The Journal of cell biology.

[367]  Mohini Jangi,et al.  Functional Surfaces on the Actin-binding Protein Coronin Revealed by Systematic Mutagenesis* , 2010, The Journal of Biological Chemistry.

[368]  D. Drubin,et al.  Loss of Aip1 reveals a role in maintaining the actin monomer pool and an in vivo oligomer assembly pathway , 2010, The Journal of cell biology.

[369]  T. Pollard,et al.  Crystal Structure of Arp2/3 Complex , 2001, Science.

[370]  Y. Ho,et al.  Characterization of the yeast amphiphysins Rvs161p and Rvs167p reveals roles for the Rvs heterodimer in vivo. , 2005, Molecular biology of the cell.

[371]  P. Lappalainen,et al.  Mammalian twinfilin sequesters ADP‐G‐actin and caps filament barbed ends: implications in motility , 2006, The EMBO journal.

[372]  A. Bretscher Fimbrin is a cytoskeletal protein that crosslinks F-actin in vitro. , 1981, Proceedings of the National Academy of Sciences of the United States of America.

[373]  S. Almo,et al.  In Vivo Importance of Actin Nucleotide Exchange Catalyzed by Profilin , 2000, The Journal of cell biology.

[374]  M. Wilmanns,et al.  Structural Conservation between the Actin Monomer-binding Sites of Twinfilin and Actin-depolymerizing Factor (ADF)/Cofilin* , 2002, The Journal of Biological Chemistry.

[375]  J. Rowley,et al.  The t(10;11)(p13;q14) in the U937 cell line results in the fusion of the AF10 gene and CALM, encoding a new member of the AP-3 clathrin assembly protein family. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[376]  B. Wendland Epsins: adaptors in endocytosis? , 2002, Nature Reviews Molecular Cell Biology.

[377]  P. Rubenstein,et al.  Acceleration of Yeast Actin Polymerization by Yeast Arp2/3 Complex Does Not Require an Arp2/3-activating Protein* , 2005, Journal of Biological Chemistry.

[378]  Gary G. Borisy,et al.  Arp2/3 Complex and Actin Depolymerizing Factor/Cofilin in Dendritic Organization and Treadmilling of Actin Filament Array in Lamellipodia , 1999, The Journal of cell biology.

[379]  H. Kueh,et al.  Rapid actin monomer–insensitive depolymerization of Listeria actin comet tails by cofilin, coronin, and Aip1 , 2006, The Journal of cell biology.

[380]  P. Matsudaira,et al.  F-actin binding and bundling properties of fimbrin, a major cytoskeletal protein of microvillus core filaments. , 1981, The Journal of biological chemistry.

[381]  L. Blanchoin,et al.  Identification of Arabidopsis cyclase-associated protein 1 as the first nucleotide exchange factor for plant actin. , 2007, Molecular biology of the cell.

[382]  S. Emr,et al.  Regulation of PI4,5P2 synthesis by nuclear–cytoplasmic shuttling of the Mss4 lipid kinase , 2003, The EMBO journal.

[383]  K. Ayscough,et al.  Yeast Endocytic Adaptor AP-2 Binds the Stress Sensor Mid2 and Functions in Polarized Cell Responses , 2014, Traffic.

[384]  R. Loewith,et al.  Functional interactions between sphingolipids and sterols in biological membranes regulating cell physiology. , 2009, Molecular biology of the cell.

[385]  T D Pollard,et al.  Molecular mechanisms controlling actin filament dynamics in nonmuscle cells. , 2000, Annual review of biophysics and biomolecular structure.

[386]  B. Haarer,et al.  Purification of profilin from Saccharomyces cerevisiae and analysis of profilin-deficient cells , 1990, The Journal of cell biology.

[387]  David K Wilson,et al.  The Structure of Aip1p, a WD Repeat Protein That Regulates Cofilin-mediated Actin Depolymerization* , 2003, Journal of Biological Chemistry.

[388]  Å. Engqvist-Goldstein,et al.  Association of mouse actin-binding protein 1 (mAbp1/SH3P7), an Src kinase target, with dynamic regions of the cortical actin cytoskeleton in response to Rac1 activation. , 2000, Molecular biology of the cell.

[389]  T. Pollard,et al.  Cofilin Dissociates Arp2/3 Complex and Branches from Actin Filaments , 2009, Current Biology.

[390]  H. Pelham,et al.  Arrestin-Mediated Endocytosis of Yeast Plasma Membrane Transporters , 2009, Traffic.

[391]  I. Gamache,et al.  The Spermidine Transport System Is Regulated by Ligand Inactivation, Endocytosis, and by the Npr1p Ser/Thr Protein Kinase in Saccharomyces cerevisiae * , 1998, The Journal of Biological Chemistry.

[392]  Kenneth A. Taylor,et al.  Structural Organization of the Actin Cytoskeleton at Sites of Clathrin-Mediated Endocytosis , 2011, Current Biology.

[393]  Beverly Wendland,et al.  Regulators of yeast endocytosis identified by systematic quantitative analysis , 2009, The Journal of cell biology.

[394]  R. Lipowsky,et al.  Individual Actin Filaments in a Microfluidic Flow Reveal the Mechanism of ATP Hydrolysis and Give Insight Into the Properties of Profilin , 2011, PLoS biology.

[395]  M. Balasubramanian,et al.  In vitro contraction of cytokinetic ring depends on myosin II but not on actin dynamics , 2013, Nature Cell Biology.

[396]  B. Appleton,et al.  The crystal structure of murine coronin-1: a regulator of actin cytoskeletal dynamics in lymphocytes. , 2006, Structure.

[397]  B. André,et al.  The Npr1 Kinase Controls Biosynthetic and Endocytic Sorting of the Yeast Gap1 Permease* , 2001, The Journal of Biological Chemistry.

[398]  K. Ayscough,et al.  Yeast Dynamin Vps1 and Amphiphysin Rvs167 Function Together During Endocytosis , 2012, Traffic.

[399]  Adam C. Martin,et al.  Negative regulation of yeast Eps15-like Arp2/3 complex activator, Pan1p, by the Hip1R-related protein, Sla2p, during endocytosis. , 2006, Molecular biology of the cell.

[400]  John A.G. Briggs,et al.  Plasma Membrane Reshaping during Endocytosis Is Revealed by Time-Resolved Electron Tomography , 2012, Cell.

[401]  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.

[402]  D. Drubin,et al.  A conserved proline-rich region of the Saccharomyces cerevisiae cyclase-associated protein binds SH3 domains and modulates cytoskeletal localization , 1996, Molecular and cellular biology.

[403]  T. Holak,et al.  Lifeact: a versatile marker to visualize F-actin , 2008, Nature Methods.

[404]  G. Sprague,,et al.  Clathrin facilitates the internalization of seven transmembrane segment receptors for mating pheromones in yeast , 1993, The Journal of cell biology.

[405]  Emanuele Cocucci,et al.  Distinct Dynamics of Endocytic Clathrin-Coated Pits and Coated Plaques , 2009, PLoS biology.

[406]  R. Dominguez,et al.  Glia Maturation Factor (GMF) Interacts with Arp2/3 Complex in a Nucleotide State-dependent Manner* , 2013, The Journal of Biological Chemistry.

[407]  S. Ono,et al.  Microscopic Evidence That Actin-interacting Protein 1 Actively Disassembles Actin-depolymerizing Factor/Cofilin-bound Actin Filaments* , 2004, Journal of Biological Chemistry.

[408]  H. Riezman,et al.  The END3 gene encodes a protein that is required for the internalization step of endocytosis and for actin cytoskeleton organization in yeast , 1994, Molecular biology of the cell.

[409]  Zooming in on the molecular mechanisms of endocytic budding by time-resolved electron microscopy , 2014, Cellular and Molecular Life Sciences.

[410]  Y. Hayashizaki,et al.  NMR solution structures of actin depolymerizing factor homology domains , 2009, Protein science : a publication of the Protein Society.

[411]  T. Mitchison,et al.  Interaction of human Arp2/3 complex and the Listeria monocytogenes ActA protein in actin filament nucleation. , 1998, Science.

[412]  S. Schmid,et al.  Actin Assembly Plays a Variable, but not Obligatory Role in Receptor‐Mediated Endocytosis , 2000, Traffic.

[413]  S. Almo,et al.  Structure of the actin crosslinking core of fimbrin. , 2004, Structure.

[414]  L. Castagnoli,et al.  Intersectin, a Novel Adaptor Protein with Two Eps15 Homology and Five Src Homology 3 Domains* , 1998, The Journal of Biological Chemistry.

[415]  M. Schuldiner,et al.  A novel single-cell screening platform reveals proteome plasticity during yeast stress responses , 2013, The Journal of cell biology.

[416]  B. André,et al.  The yeast Npi1/Rsp5 ubiquitin ligase lacking its N-terminal C2 domain is competent for ubiquitination but not for subsequent endocytosis of the gap1 permease. , 1999, Biochemical and biophysical research communications.

[417]  S. Narumiya,et al.  An Order of Magnitude Faster AIP1-Associated Actin Disruption than Nucleation by the Arp2/3 Complex in Lamellipodia , 2009, PloS one.

[418]  D. Sabatini,et al.  Actin microfilaments play a critical role in endocytosis at the apical but not the basolateral surface of polarized epithelial cells , 1993, The Journal of cell biology.

[419]  T. Pollard,et al.  Cellular Motility Driven by Assembly and Disassembly of Actin Filaments , 2003, Cell.

[420]  Brian J. Stevenson,et al.  The WASp homologue Las17p functions with the WIP homologue End5p/verprolin and is essential for endocytosis in yeast , 1998, Current Biology.

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

[422]  Howard Riezman,et al.  Ubiquitination of a Yeast Plasma Membrane Receptor Signals Its Ligand-Stimulated Endocytosis , 1996, Cell.

[423]  T. Pollard,et al.  Review of the mechanism of processive actin filament elongation by formins. , 2009, Cell motility and the cytoskeleton.

[424]  Jonathan A. Cooper,et al.  Dynamics of capping protein and actin assembly in vitro: uncapping barbed ends by polyphosphoinositides , 1996, The Journal of cell biology.

[425]  D. Amberg,et al.  Three-dimensional imaging of the yeast actin cytoskeleton through the budding cell cycle. , 1998, Molecular biology of the cell.

[426]  B. Andrews,et al.  Clathrin light chain directs endocytosis by influencing the binding of the yeast Hip1R homologue, Sla2, to F-actin , 2011, Molecular biology of the cell.