Contents


Symposium Participants    v

Foreword    xix



Protein Translocation in the Endoplasmic Reticulum and Golgi

Unidirectional and Bidirectional Protein Traffic across Membranes G. Blobel    1

Coat Proteins and Selective Protein Packaging into Transport Vesicles R. Schekman, C. Barlowe, S. Bednarek, J. Campbell, T. Doering, R. Duden, M. Kuehn, M. Rexach, T. Yeung, and L. Orci    11

Targeting Pathways to the Endoplasmic Reticulum Membrane J.D. Brown, D.T.W. Ng, S.C. Ogg, and P. Walter    23

Posttranslational Protein Transport into the Endoplasmic Reticulum S. Panzner, L. Dreier, E. Hartmann, S. Kostka, and T.A. Rapoport    31

Protein Insertion into the Membrane of the Endoplasmic Reticulum: The Architecture of the Translocation Site B. Martoglio and B. Dobberstein    41

Nascent-polypeptide-associated Complex: A Bridge between Ribosome and Cytosol B. Lauring, S. Wang, H. Sakai, T.A. Davis, B. Wiedmann, G. Kreibich, and M. Wied- Mann    47

Protein-conducting Channels for the Translocation of Proteins into and across Membranes S.M. Simon    57

The Environment of Nascent Secretory and Membrane Proteins at the Endoplasmic Reticulum Membrane during Translocation and Integration A.E. Johnson, S. Liao, J. Lin, B. Hamman, H. Do, A. Cowie, and D.W. Andrews    71

The Saccharomyces cerevisiae Oligosaccharyltransferase: A Large Hetero-oligomeric Complex in the Endoplasmic Reticulum D. Karaoglu, S. Silberstein, D.J. Kelleher, and R. Gilmore    83

The Erythropoietin Receptor: Biogenesis, Dimerization, and Intracellular Signal Transduction H.F. Lodish, D.J. Hilton, U. Klingmüller, S.S. Watowich, and H. Wu    93

SNAREs Involved in Traffic through the Golgi Complex H.R.B Pelham, D.K. Banfield, and M.J. Lewis    105

Targeting to the Endoplasmic Reticulum by Dilysine Motifs: Role of Coat Proteins P. Cosson and F. Letourneur    113

Factors Mediating the Late Stages of ER-to-Golgi Transport in Yeast Y. Jiang, M. Sacher, B. Singer-Krüger, J.P. Lian, S. Stone, and S. Ferro-Novick    119

Regulated Export of Cargo from the Endoplasmic Reticulum of Mammalian Cells S. Bannykh, M. Aridor, H. Plutner, T. Rowe, and W.E. Balch    127

Molecules and Vesicle Coats Involved in the Budding of Exocytotic Vesicles from the trans-Golgi Network J. Wang, M.S. Ladinsky, and K.E. Howell    139

Mapping a Retention Signal for Golgi Localization of a Viral Spike Protein Complex R.F. Pettersson, A. Andersson, and L. Melin    147

Novel Protein Kinase/Phosphatidylinositol 3-Kinase Complex Essential for Receptormediated Protein Sorting to the Vacuole in Yeast J.H. Stack, P.K. Herman, D.B. DeWald, E.G. Marcusson, J. Lin Cereghino, B.F. Horazdovsky, and S.D. Emr    157



Protein Trafficking at the Cell Surface

Control of Exocytosis in Yeast P. Novick, M.D. Garrett, P. Brennwald, A. Lauring, F.P. Finger, R. Collins, and D.R. TerBush    171

Regulation of Post-Golgi Vesicle Production in an In Vitro System J.-P. Simon, I.E. Ivanov, M. Ren, J. Zeng, B. Shopsin, D. Hersh, P. Tempst, H. Erdjument-Bromage, M. Lui, C. De Lemos-Chiarandini, M. Rosenfeld, D. Gravotta, T. Morimoto, M. Adesnik, and D.D. Sabatini    179

Late ATP-dependent and Ca++ -activated Steps of Dense Core Granule Exocytosis T.F.J. Martin, J.C. Hay, A. Banerjee, V.A. Barry, K. Ann, H.-C. Yom, B.W. Porter, and J.A. Kowalchyk    197

Membrane Transport from Early to Late Endosomes F. Aniento and J. Gruenberg    205

The GDP/GTP Cycle of Rab5 in the Regulation of Endocytotic Membrane Traffic G. Vitale, K. Alexandrov, O. Ullrich, H. Horiuchi, A. Giner, C. Dobson, O. Baykova, H. Gournier, H. Stenmark, and M. Zerial    211

Selective Membrane Recruitment of Rab GTPases S.R. Pfeffer, T. Soldati, H. Geissler, C. Rancaño, and B. Dirac-Svejstrup    221

The ARF GTPases: Defining Roles in Membrane Traffic and Organelle Structure J.G. Donaldson, H. Radhakrishna, and P.J. Peters     229

Role of Dynamin in Clathrin-coated Vesicle Formation T. Baba, H. Damke, J.E. Hinshaw, K. Ikeda, S.L. Schmid, and D.E. Warnock     235

Inhibitors That Block Both the Internalization of Caveolae and the Return of Plasmalemmal Vesicles E.J. Smart, K. Estes, and R.G.W. Anderson    243

Assembly and Intracellular Transport of MHC Class I and Class II Molecules M.R. Jackson, K. Früh, L. Karlsson, L. Teyton, Y. Yang, and P.A. Peterson     249

Trafficking and Assembly of MHC Molecules: How Viruses Elude the Immune System H.L. Ploegh     263

Assembly, Peptide Loading, and Transport of MHC Class I Molecules in a Calnexin- negative Cell Line B.K. Sadasivan, A. Cariappa, G.L. Waneck, and P. Cresswell    267

Essential Features of the Pathway for Protein Translocation across the Escherichia coli Cytoplasmic Membrane C. Murphy, W. Prinz, M. Pohlschrîder, A. Derman, and J. Beckwith    277

On the Road to Translocase W. Wickner, M.R. Leonard, and A. Economou    285

Sequence Comparison of Yeast ATP-binding Cassette Proteins S. Michaelis and C. Berkower    291



Synaptic Trafficking

A Systematic Approach to Studying Synaptic Function in Vertebrates R. Janz and T.C. Südhof     309

Biogenesis of Neurosecretory Vesicles W.B. Huttner, M. Ohashi, R.H. Kehlenbach, F.A. Barr, R. Bauerfeind, O. Bräunling, D. Corbeil, M. Hannah, H.A. Pasolli, A. Schmidt, A.A. Schmidt, C. Thiele, Y. Wang, A. Krîmer, and H.-H. Gerdes    315

Botulinum and Tetanus Neurotoxins: Emerging Tools for the Study of Membrane Fusion R. Jahn, P.I. Hanson, H. Otto, and G. Ahnert-Hilger    329

Proteins Involved in Synaptic Vesicle Docking and Fusion M.E. Burns, S.A. Beushausen, G.J. Chin, D. Tang, W.M. DeBello, T. Dresbach, V. O'Connor, F.E. Schweizer, S.S.-H. Wang, S.W. Whiteheart, L.A. Hawkey, H. Betz, and G.J. Augustine    337

Calcium-dependent Vesicular Exocytosis: From Constitutive to Regulated Secretion M. Poo, Y. Dan, H. Song, T. Morimoto, and S. Popov    349

Insights into the Molecular Mechanisms of Synaptic Vesicle Docking and Fusion A.E. Ting and R.H. Scheller     361

Drosophila Genetics and the Functions of Synaptic Proteins K. Parfitt, N. Reist, J. Li, R. Burgess, D. Deitcher, A. DiAntonio, and T.L. Schwarz    371

Recycling of Synaptic Vesicle Membrane Components Y. Kaneda, E. Grote, F. Bonzelius, C. Desnos, G. Herman, L. Clift-O'Grady, and R.B. Kelly    379

The Last Seconds in the Life of a Secretory Vesicle T.D. Parsons, J.R. Coorssen, H. Horstmann, A.K. Lee, F.W. Tse, and W. Almers    389

Molecular Mechanisms in Synaptic Vesicle Endocytosis R. Bauerfeind, C. David, T. Galli, P.S. McPherson, K. Takei, and P. De Camilli    397



Protein Folding

Calnexin, Calreticulin, and Bip/Kar2p in Protein Folding D.N. Hebert, J.F. Simons, J.R. Peterson, and A. Helenius    405

Binding Sites for Hsp70 Molecular Chaperones in Natural Proteins M.-J. Gething, S. Blond-Elguindi, J. Buchner, A. Fourie, G. Knarr, S. Modrow, L. Nanu, M. Segal, and J. Sambrook    417

Mechanisms and Pathways of Chaperone-mediated Protein Folding F.U. Hartl    429

Kinesis of Polypeptide during GroEL-mediated Folding A.L. Horwich, J.S. Weissman, and W.A. Fenton    435

Complex Multigene Family of Functionally Distinct Hsp70s of Yeast E. Craig, T. Ziegelhoffer, J. Nelson, S. Laloraya, and J. Halladay    441

The Role of Hsp104 in Stress Tolerance and [PSI+] Propagation in Saccharomyces cerevisiae S. Lindquist, M.M. Patino, Y.O. Chernoff, A.S. Kowal, M.A. Singer, S.W. Liebman, K.-H. Lee, and T. Blake    451



Protein Degradation

The N-end Rule A. Varshavsky    461

Functions of the Proteasome in Antigen Presentation A.L. Goldberg, M. Gaczynska, E. Grant, M. Michalek, and K.L. Rock    479

The Ubiquitin-mediated Proteolytic System: Involvement of Molecular Chaperones, Degradation of Oncoproteins, and Activation of Transcriptional Regulators A. Ciechanover, A. Laszlo, B. Bercovich, I. Stancovski, I. Alkalay, Y. Ben-Neriah, and A. Orian    491

The DOA Pathway: Studies on the Functions and Mechanisms of Ubiquitin-dependent Protein Degradation in the Yeast Saccharomyces cerevisiae M. Hochstrasser, F.R. Papa, P. Chen, S. Swaminathan, P. Johnson, L. Stillman, A.Y. Amerik, and S.-J. Li    503

Structure and Function of the 20S Proteasome and of Its Regulatory Complexes A. Lupas, P. Zwickl, T. Wenzel, E. Seemüller, and W. Baumeister    515

Catalytic Mechanism of the 20S Proteasome of Thermoplasma acidophilum Revealed by X-ray Crystallography D. Stock, L. Ditzel, W. Baumeister, R. Huber, and J. Löwe     525

Selective, Energy-dependent Proteolysis in Escherichia coli S. Gottesman, S. Wickner, Y. Jubete, S.K. Singh, M. Kessel, and M. Maurizi    533



Membrane Dynamics

Mitotic Disassembly and Reassembly of the Golgi Apparatus T.P. Levine, T. Misteli, C. Rabouille, and G. Warren    549

Reconstitution of Golgi Stacks from Vesiculated Golgi Membranes in Permeabilized Cells U. Acharya and V. Malhotra    559

Regulation of Endocytosis, Exocytosis, and Shape by Membrane Tension J. Dai and M.P. Sheetz    567

Membrane Fusion by Influenza Hemagglutinin J.J. Skehel, T. Bizebard, P.A. Bullough, F.M. Hughson, M. Knossow, D.A. Steinhauer, S.A. Wharton, and D.C. Wiley    573

Membrane Fusion: The Influenza Paradigm J.M. White    581

Intermediates in Membrane Fusion J. Zimmerberg, S.S. Vogel, T. Whalley, I. Plonsky, A. Sokoloff, A. Chanturia, and L.V. Chernomordik    589

Role of Water in Short-range Repulsive and Attractive Interactions between Macromolecules and Bilayers S.A. Simon and T.J. McIntosh    601



Protein Import into Organelles

Import and Folding of Proteins by Mitochondria T. Lithgow, M. Horst, S. Rospert, A. Matouschek, V. Haucke, and G. Schatz    609

Dissection of Protein Translocation across the Mitochondrial Outer and Inner Membranes M. Brunner, H.-C. Schneider, R. Lill, and W. Neupert    619

Molecular Dissection of the Mechanism of Protein Import into Chloroplasts F. Tian, Y. Ma, A. Kouranov, S.E. LaSala, and D.J. Schnell    629

Characterization of the Yeast and Human Receptors for the Carboxy-terminal Tripeptide Peroxisomal Targeting Signal E.A.C. Wiemer, S.R. Terlecky, W.M. Nuttley, and S. Subramani    637

Transport of Proteins and Metabolites across the Impermeable Membrane of Peroxisomes H.F. Tabak, Y. Elgersma, E. Hettema, M.M. Franse, T. Voorn- Brouwer, and B. Distel    649

Protein Import into Peroxisomes: An Exception to the Rule? B. Huhse and W.-H. Kunau    657



Nuclear Trafficking

Signal Sequences That Target Nuclear Import and Nuclear Export of Pre-mRNA- binding Proteins W.M. Michael, H. Siomi, M. Choi, S. Piñol-Roma, S. Nakielny, Q. Liu, and G. Dreyfuss    663

Nuclear Localization of a Cap-binding Protein Complex E. Izaurralde, C. McGuigan, and I.W. Mattaj    669

Nucleocytoplasmic Trafficking: What Role for Repeated Motifs in Nucleoporins? E. Fabre, N.L. Schlaich, and E.C. Hurt    677

Cytoplasmic Factors in NLS-mediated Targeting to the Nuclear Pore Complex S.A. Adam, E.J.H. Adam, N.C. Chi, and G.D. Visser    687

Roles of Importin in Nuclear Protein Import D. Görlich and R.A. Laskey    695

Soluble Factors Required for Nuclear Protein Import M.S. Moore and G. Blobel    701

Biochemical and Structural Analysis of Nuclear Protein Import F. Melchior, T. Guan, B. Paschal, and L. Gerace    707



Cell Polarity

Programming of Cell Polarity in Budding Yeast by Endogenous and Exogenous Signals I. Herskowitz, H.-O. Park, S. Sanders, N. Valtz, and M. Peter    717

Establishment of Cell Polarity in Yeast J.R. Pringle, E. Bi, H.A. Harkins, J.E. Zahner, C. De Virgilio, J. Chant, K. Corrado, and H. Fares    729

Molecular Sorting of Membrane Proteins in Polarized and Nonpolarized Cells I. Mellman    745

Dissecting the Molecular Mechanisms of Polarized Membrane Traffic: Reconstitution of Three Transport Steps in Epithelial Cells Using Streptolysin-O Permeabilization F. Lafont, K. Simons, and E. Ikonen    753

Generation of Epithelial Cell Polarity: Roles for Protein Trafficking, Membrane- Cytoskeleton, and E-Cadherin-mediated Cell Adhesion R.W. Mays, W.J. Nelson, and J.A. Marrs    763

Regulation of Protein Traffic in Polarized Epithelial Cells: The Polymeric Immunoglobulin Receptor Model K.E. Mostov, Y. Altschuler, S.J. Chapin, C. Enrich, S.-H. Low, F. Luton, J. Richman-Eisenstat, K.L. Singer, K. Tang, and T. Weimbs    775



Molecular Motors

Myosin Structure and Function J.A. Spudich, J. Finer, B. Simmons, K. Ruppel, B. Patterson, and T. Uyeda    783

Statistical Kinetics of Processive Enzymes M.J. Schnitzer and S.M. Block    793

Targeting of Cytoplasmic Dynein to Membranous Organelles and Kinetochores via Dynactin R.B. Vallee, K.T. Vaughan, and C.J. Echeverri    803

Analysis of Nuclear Migration in Aspergillus nidulans X. Xiang, A.H. Osmani, S.A. Osmani, C.H. Roghi, D.A. Willins, S. Beckwith, G. Goldman, Y. Chiu, M. Xin, B. Liu, and N.R. Morris    813

Summary: G.E. Palade    821