On Quantitative Biology Volume Lx

COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY

VOLUME LX COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY

VOLUME LX

Protein Kinesis: The Dynamics of Protein Trafficking and Stability

COLD SPRING HARBOR LABORATORY PRESS 1995 COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY VOLUME LX

9 by Cold Spring Harbor Laboratory Press International Standard Book Number 0-87969-069-0 (cloth) International Standard Book Number 0-87969-070-4 (paper) International Standard Serial Number 0091-7451 Library of Congress Catalog Card Number 34-8174 Printed in the United States of America All rights reserved

COLD SPRING HARBOR SYMPOSIA ON QUANTITATIVE BIOLOGY Founded in 1933 by REGINALD G. HARRIS Director of the Biological Laboratory 1924 to 1936

Previous Symposia Volumes I (1933) Surface Phenomena XXIX (1964) Human Genetics II (1934) Aspects of Growth XXX (1965) Sensory Receptors III (1935) Photochemical Reactions XXXI (1966) The Genetic Code IV (1936) Excitation Phenomena XXXII (1967) Antibodies V (1937) Internal Secretions XXXIII (1968) Replication of DNA in Microorganisms VI (1938) Protein Chemistry XXXIV (1969) The Mechanism of Protein Synthesis VII (1939) Biological Oxidations XXXV (1970) Transcription of Genetic Material VIII (1940) Permeability and the Nature of Cell Membranes XXXVI (1971) Structure and Function of Proteins at the IX (1941) Genes and Chromosomes: Structure and Organiza- Three-dimensional Level tion XXXVII (1972) The Mechanism of Muscle Contraction X (1942) The Relation of Hormones to Development XXXVIII (1973) Chromosome Structure and Function XI (1946) Heredity and Variation in Microorganisms XXXIX (1974) Tumor Viruses XII (1947) Nucleic Acids and Nucleoproteins XL (1975) The Synapse XIII (1948) Biological Applications of Tracer Elements XLI (1976) Origins of Lymphocyte Diversity XIV (1949) Amino Acids and Proteins XLII (1977) Chromatin XV (1950) Origin and Evolution of Man XLIII (1978) DNA: Replication and Recombination XVI (1951) Genes and Mutations XLIV (1979) Viral Oncogenes XVII (1952) The Neuron XLV (1980) Movable Genetic Elements XVIII (1953) Viruses XLVI (1981) Organization of the Cytoplasm XIX (1954) The Mammalian Fetus: Physiological Aspects of XLVII (1982) Structures of DNA Development XLVIII (1983) Molecular Neurobiology XX (1955) Population Genetics: The Nature and Causes of XLIX (1984) Recombination at the DNA Level Genetic Variability in Population L (1985) Molecular Biology of Development XXI (1956) Genetic Mechanisms: Structure and Function LI (1986) Molecular Biology of Homo sapiens XXII (1957) Population Studies: Animal Ecology and De- LII (1987) Evolution of Catalytic Function mography LIII (1988) Molecular Biology of Signal Transduction XXIII (1958) Exchange of Genetic Material: Mechanism and LIV (1989) Immunological Recognition Consequences LV (1990) The Brain XXIV (1959) Genetics and Twentieth Century Darwinism LVI (1991)The Cell Cycle XXV (1960) Biological Clocks LVII (1992) The Cell Surface XXVI (1961) Cellular Regulatory Mechanisms LVIII (1993) DNA and Chromosomes XXVII (1962) Basic Mechanisms in Animal Virus Biology LIX (1994) The Molecular Genetics of Cancer XXVIII (1963) Synthesis and Structure of Macromolecules

Front Cover (Paperback): 14-fold averaged difference electron density showing seven LLnL inhibitors sitting around the central cavity formed by 13 subunits. (For details, see D. Stock et al., p. 528, this volume.) Back Cover (Paperback): Schematic representation of possible topological organization of cytoplasmic dynein and dynactin in the prometaphase kinetochore. (For details, see R.B. Vallee et al., p. 809, this volume.) Authorization to photocopy items for internal or personal use, or the internal or personal use of specific clients, is granted by Cold Spring Harbor Laboratory for libraries and other users registered with the Copyright Clearance Center (CCC) Transactional Reporting Service, provided that the base fee of $5.00 per article is paid directly to CCC, 222 Rosewood Dr., Danvers, MA 01923. [0-87969-069-0/95 $5.00 + .00]. This consent does not extend to other kinds of copying, such as copying for general distribution, for advertising or promotional purposes, for creat- ing new collective works, or for resale. All Cold Spring Harbor Laboratory Press publications may be ordered directly from Cold Spring Harbor Laboratory Press, 10 Skyline Drive, Plainview, NY 11803. Phone: 1-800-843-4388 in Continental U.S. and Canada. All other locations (516) 349-1930. FAX: (516) 349-1946. E-mail: [email protected]. For a complete catalog of all Cold Spring Harbor Laboratory Press publications, visit our World Wide Web Site http://www.cshl.org/ Symposium Participants

ABRAMS, JOAN, Dept. of Physiology, Tufts University, BAUMEISTER, WOLFGANG, Dept. of Structural Biol- Boston, Massachusetts ogy, Max-Planck Institute for Biochemistry, ADAM, STEPHEN, Dept. of Cell and Molecular Biology, Martinsried Bavaria, Germany Northwestern University Medical School, Chicago, BAZINET, CHRISTOPHER, Dept. of Biological Sciences, Illinois St. John's University, Jamaica, New York ADESNIK, MILTON, Dept. of Cell Biology, New York BECKWITH, JONATHAN, Dept. of Microbiology and University Medical Center, New York Molecular Genetics, Harvard Medical School, Bos- AHN, KWANGSEOG, Dept. of Immunology, R.W. ton, Massachusetts Johnson Pharmaceutical Research Institute, La BEN-NERIAH, YINON, Dept. of Immunology, Hebrew Jolla, California University Hadassah Medical School, Lautenberg AITCHISON, JOHN, Dept. of Cell Biology, Rockefeller Center, Jerusalem, Israel University, New York, New York BERMAN, PHILIP, Dept. of Immunology, Genentech, ALBANESI,JOSEPH, Dept. of Pharmacology, University Inc., South San Francisco, California of Texas Southwestern Medical Center, Dallas BERNSTEIN, HARRIS, Genetics and Biochemistry ALLEN, SIMON, Dept. of Biochemistry, School of Branch/NIDDK, National Institutes of Health, Biological Sciences, University of Manchester, Bethesda, Maryland United Kingdom BICKEL, PERRY, Dept. of Biology, Whitehead Institute ALMERS, WOLFHARD, Max-Planck Instittit ftir for Biomedical Research, Cambridge, Mas.sachu- Medizinische Forschung, Heidelberg, Germany setts ANDERSON, RICHARD, Dept. of Cell Biology and BIJLMAKERS, MARIE-JOSE, Dept. of Molecular Cell Neuroscience, University of Texas Southwestern Biology, MRC Laboratory, London, United King- Medical Center, Dallas dom ANDERSSON, AGNETA, Ludwig Institute for Cancer BLOBEL, GUNTER, Dept. of Cell Biology, Rockefeller Research, Stockholm, Sweden University, New York, New York ANTON, LUIS, Viral Immunology Section, National In- BLOCK, STEVEN, Dept. of Molecular Biology, Prince- stitute of Allergy and Infectious Diseases, National ton University, Princeton, New Jersey Institutes of Health, Bethesda, Maryland BLOOM, THEODORA, Current Biology, London, United ARMSTRONG, JOHN, Dept. of Cellular Science, Glaxo Kingdom Group Research Limited, Middlesex, United King- BOBIEH, JOSEPH, Dept. of Chemistry, Texas Christian dom University, Fort Worth ARNESON, LYNNE, Dept. of Molecular Genetics and BOER, JUDITH, Dept. of Genetics, St. Jude Children's Cell Biology, University of Chicago, Illinois Research Hospital, Memphis, Tennessee AUGENSTEIN, SCOTT, Dynal, Inc., Lake Success, New BOMSEL, MORGANE, Dept. of Cell Biology, Institut York Cochin de Genetique Moleculaire, Paris, France AUGUSTINE, GEORGE, Dept. of Neurobiology, Duke BROOKS, DOUG, Dept. of Chemical Pathology, University, Durham, North Carolina Women's and Children's Hospital, North AXELROD, AMY, Cell, Cambridge, Massachusetts Adelaide, South Australia BABST, MARKUS, ETH Zurich, Mikrobiologisches In- BROSTEDT, ERICA, Cellular Biochemistry and stitut, Zurich, Switzerland Biophysics Program, Memorial Sloan-Kettering BACHER, GERALD, Zentrum fur Molekulare Biologie, Cancer Research Institute, New York, New York University of Heidelberg, Germany BROWNAWELL, AMY, Dept. of Pharmacology, Univer- BALCH, WILLIAM, Dept. of Molecular Biology, The sity of Virginia Health Sciences Center, Char- Scripps Research Institute, La Jolla, California lottesville BARBOSA, MARIA, Dept. of Medicine, Division of BRUNETTI,CRAIG, Dept. of Pathology, McMaster Uni- Rheumatology, University of Florida, Gainesville versity, Hamilton, Ontario, Canada BARYLKO, BARBARA,Dept. of Pharmacology, Univer- BRUNNER, MICHAEL, Physiologische Chemie, Univer- sity of Texas Southwestern Medical Center, Dallas sit,it Munchen, Munich, Germany vi SYMPOSIUM PARTICIPANTS

BURGESS, TERESA, Dept. of Mammalian Cell CREVEL, ISABELLE,Molecular Motors Group, Marie Molecular Biology, Amgen Incorporated, Thou- Curie Research Institute, The Chart, United King- sand Oaks, California dom BURKHARDT, JANIS, Cell Biology Programme, Euro- DARGENT, BENED[CTE, Neurobiologie Des Canaux pean Molecular Biology Laboratory, Heidelberg, Ioniques, INSERM, Marseille, France Germany DARNELL, ROBERT, Rockefeller University, New BURTON, JANET, Dept. of Cell Biology, Yale School of York, New York Medicine, New Haven, Connecticut DAVEY, JOHN, School of Biochemistry, University of CAKY, MYRON, Dept. of Neurology, McGill Univer- Birmingham, United Kingdom sity/Montreal Neurological Institute, Quebec, Can- DAVILA-APONTE, JENNY, Plant Research Laboratory, ada Michigan State University, East Lansing CASANOVA, JAMES, Pediatric GI, Massachusetts Gen- DAVIS, ELAINE, Dept. of Cell Biology and Physiology, eral Hospital East, Charlestown Washington University School of Medicine, St. CASSEL, DAN, Dept. of Biology, Technion, Haifa, Is- Louis, Missouri rael DAVIS, NICK, Dept. of Pharmacology and Surgery, CATELLI, MARIA-GRAZIA,Hormones Laboratory, IN- Wayne State University School of Medicine, De- SERM, Kremlin Bicetre, France troit, Michigan CERUNDOLO, VINCENZO, Dept. of Molecular Im- DE CAMILLI,PIETRO, Dept. of Cell Biology, Yale Uni- munology, Institute of Molecular Medicine, Uni- versity School of Medicine, New Haven, Con- versity of Oxford, United Kingdom necticut CHANG, HENRY, Dept. of Molecular and Cell Biology, DE LEMOS-CHIARANDINI,CARMEN, Dept. of Cell Biol- University of California, Berkeley ogy, New York University School of Medicine, CHEN, YIH-TAI, Dept. of Molecular and Cellular New York Physiology, Stanford University School of Medi- DE SOUZA, NATALIE,Dept. of Cell Biology, Rockefel- cine, Stanford, California ler University, New York, New York CHEUNG, PETER, Dept. of Pathology, Cancer Research DEMPSEY, PETER, Dept. of Medicine and Ceil Biology, Group, McMaster University, Health Science Vanderbilt University, Nashville, Tennessee Centre, Hamilton, Ontario, Canada DERETIC, DUSANKA, Dept. of Pathology, University of CHO, CHARLES, Dept. of Chemistry, Schultz Research Texas Health Sciences Center, San Antonio Group, University of California, Berkeley DICKEY, BURTON, Dept. of Medicine and Cell Biology, CIECHANOVER, AARON, Dept. of Biochemistry, Tech- Baylor College of Medicine, Houston, Texas nion, Israel Institute of Technology, Bat Galim, DOBBERSTEIN, BERNHARD, Dept. of Molecular Biol- Haifa, Israel ogy, University of Heidelberg, Germany CLAIRMONT, KEVIN, Dept. of Metabolic Disorders Re- DONALDSON, JULIE, Cell Biology and Metabolism search, Bayer, Inc., West Haven, Connecticut Branch, National Institutes of Child Health and CLINE, KEN, Dept. of Horticultural Science, University Human Development, National Institutes of of Florida, Gainesville Health, Bethesda, Maryland COFFEY, ROBERT, Dept. of Medicine and Cell Biology, DREYFUS, GEORGES, Dept. of Bioenergenetics, Uni- Vanderbilt University, Nashville, Tennessee versity of Mexico, Mexico City COLLEY, ALLEN,Dept. of Biochemistry, University of DREYFUSS, GIDEON, Dept. of Biochemistry and Edinburgh, United Kingdom Biophysics, Howard Hughes Medical Institute, CONIBEAR, ELIZABETH, Dept. of Molecular Biology, University of Pennsylvania School of Medicine, University of Oregon, Eugene Philadelphia COOPER, GARY, Dept. of Pharmacology, University of ECONOMOU, ANASTASSIOS, Dept. of Biochemistry, Medicine and Dentistry of New Jersey, Rutgers Dartmouth Medical School, Hanover, New Hamp- University, Piscataway shire CORLEY, RONALD, Dept. of Microbiology, Boston EGERTON, MARK,Dept. of Infection Research, Zeneca University Medical Center, Boston, Massachusetts Pharmaceuticals, Macclesfietd, Cheshire, United CoSSON, PIERRE, Basel Institute for Immunology, Kingdom Basel, Switzerland EGUINOA, ALICIA, Dept. of Development and Signal- CRAIG, ELIZABETH,Dept. of Biomolecular Chemistry, ling, The Babraham Institute, Cambridge, United University of Wisconsin, Madison Kingdom CRESSWELL, PETER, Dept. of Immunobiology, Yale EISENBERG, EVAN, Dept. of Cell Biology, National University School of Medicine, New Haven, Con- Heart, Lung, and Blood Institute, National In- necticut stitutes of Health, Bethesda, Maryland SYMPOSIUM PARTICIPANTS vii

EMR, SCOTt, Dept. of Cellular and Molecular Medi- GETHING, MARYJANE, Dept. of Biochemistry, Univer- cine, Howard Hughes Medical Institute, University sity of Melbourne, Parkville, Victoria, Australia of California at San Diego, La Jolla GHINEA, NICOLAE,Dept. of Hormones and Reproduc- ENDO, TOSHIYA, Dept. of Chemistry, Faculty of tion, INSERM, Le Kremlin Bicetre, France Science, Nagoya University, Chikusa-ku, Nagoya, GHOSH, HARA, Dept. of Biochemistry, McMaster Uni- Japan versity, Hamilton, Ontario, Canada ENENKEL, CORDULA,Dept. of Cell Biology, Rockefel- GILMORE, REID, Dept. of Biochemistry, University of ler University, New York, New York Massachusetts Medical School, Worcester ERIKSSON,ULF, Ludwig Institute for Cancer Research, GOLDBERG, ALFRED, Dept, of Cell Biology, Harvard Stockholm, Sweden Medical School, Boston, Massachusetts FAIX, JAN, Dept. of Cell Biology, Max-Planck Institute GONZALEZ-HALPHEN,DIEGO, Dept. of Bioenergetics, for Biochemistry, Martinsried, Germany Institute of Cell Physiology, University of Mexico, FARQUHAR, MARILYN, Dept. of Cell and Molecular Mexico City Medicine, University of California at San Diego, GORBOVITSKAIA, MARIE, Unite Thyroide, INSERM, La Jolla Le Kremlin Bicetre, France FERNANDES, DANCELLA, Dept. of Lymphocyte Biol- GORLICH, DIRK, Wellcome CRC Institute, Cambridge, ogy, Dana-Farber Cancer Institute, Boston, Mass- United Kingdom achusetts GOTTESMAN, SUSAN,Dept. of Molecular Biology, Na- FERRO-NOVICK, SUSAN, Dept. of Cell Biology, tional Cancer Institute, Bethesda, Maryland Howard Hughes Medical Institute, Yale Univer- GOULD, STEVE, Dept. of Biological Chemistry, Johns sity School of Medicine, New Haven, Connecticut Hopkins University School of Medicine, Balti- FIEDLER, KLAUS, Cell Biology Program, European more, Maryland Molecular Biology Laboratory, Heidelberg, GRAHAM, TODD, Dept. of Molecular Biology, Vander- Germany bilt University, Nashville, Tennessee FINGER, FERN, Dept. of Cell Biology, Yale University GRAVOTTA, DIEGO, Dept. of Cell Biology, New York School of Medicine, New Haven, Connecticut University Medical Center, New York FISCHER VON MOLLARD, GABRIELE, Dept. of GREENALL, CAROLE,Biological Laboratory, The Uni- Molecular Biology, University of Oregon, Eugene versity of Kent, Canterbury, United Kingdom FISHER, EDWARD,Dept. of Biochemistry, Medical Col- GREENE, LOIS, Dept. of Cell Biology, National Heart, lege of Pennsylvania, Philadelphia Lung, and Blood Institute, National Institutes of FLAVELL, DAVID, Dept. of Pathology, Southampton Health, Bethesda, Maryland General Hospital, University of Southampton, GRINDSTAFF, KENT, Dept. of Molecular and Cellular United Kingdom Physiology, Stanford University, Stanford, Califor- FLEISSNER, ERWIN, Division of Science and Mathe- nia matics, Hunter College, City University of New GRITSENKO, GALINA, Dept. of Microbiology, Boston York, New York University School of Medicine, Boston, Massachu- FLEMING, JUDY, Center for Blood Research, Boston, setts Massachusetts GRUENBERG,JEAN, Dept. of Biochemistry Sciences II, FOURIE, ANNE, Dept. of Immunology, Scripps Re- University of Geneva, Switzerland search Institute, La Jolla, California GRUHLER, ALBRECHT, Institute of Physiological FROEHLICH, JOHN, MSU-DOE Plant Research Labo- Chemistry, University of Munich, Germany ratory, Michigan State University, East Lansing Guo, QIU, Dept. of Biology, Massachusetts Institute of FROSHAUER, SUZY, Dept. of Immunology and In- Technology, Cambridge fectious Diseases, Pfizer Central Research, HALLBERG, EINAR, Dept. of Biochemistry, Arrhenius- Groton, Connecticut laboratories, Stockholm University, Sweden FUJIKI, YUKIO, Dept. of Biology, Kyushu University, HAMMAN, BRIAN, Texas A&M University Health Fukuoka, Japan Science Center, College of Medicine, College Sta- GANDY, SAM, Dept. of Neurology and Neuroscience, tion Cornell University Medical College, New York, HAND, ARTHUR,Dept. of Pediatric Dentistry, Univer- New York sity of Connecticut Health Science Center, GERACE, LARRY, Dept. of Cell Biology, Scripps Re- Farmington search Institute, La Jolla, California HANOVER, JOHN, Dept. of Cell Biochemistry, National GERST, JEFFREY, Dept. of Molecular Genetics and Institutes of Health, Bethesda, Maryland Virology, Weizmann Institute of Science, Rehovot, HARRIS, CHRIS,Dept. of Molecular Biology, Princeton Israel University, Princeton, New Jersey viii SYMPOSIUM PARTICIPANTS

HARRIS, DAVID, Dept. of Cell Biology and Physiology, ITO, KOREAKI, Dept. of Cell Biology, Institute for Washington University School of Medicine, St. Virus Research, Kyoto University, Sakyo-ku, Louis, Missouri Kyoto, Japan HARRIS, SANDRA, Dept. of Molecular Biology, Prince- IVERFELDT, KERSTIN, Dept. of Neuroehemistry and ton University, Princeton, New Jersey Neurotoxicology, Stockholm University, Sweden HARSAY, EDINA, Dept. of Biochemistry, Cornell Uni- IVESSA, ERWIN N., Dept. of Molecular Genetics, versity, Ithaca, New York Biocenter, University of Vienna, Austria HARTL, F. ULRICH,Dept. of Cellular Biochemistry and JADOT, MICHEL, Chimie Physiologique, University of Biophysics, Memorial Sloan-Kettering Cancer Notre-Dame de la Paix, Namur, Belgium Center, New York, New York JAGADISH, MITTUR, Dept. of Biomoleeular Engineer- HARTMANN, ENNO, Dept. of Cell Biology, Max- ing, Institute of Industrial Technologies, Parkville, Delbriick Center for Molecular Medicine, Berlin, Australia Germany JAHN, REINHARD, Boyer Center for Molecular Medi- HATANAKA, MASAKAZU,Institute for Virus Research, cine, Howard Hughes Medical Institute, Yale Uni- Kyoto University, Sakyo-ku, Kyoto, Japan versity School of Medicine, New Haven, Con- HAWIGER, JACEK, Dept. of Microbiology, Vanderbilt necticut University, Nashville, Tennessee JEN, CHAUYINGJACK, Dept. of Physiology, College of HEILKER, RALF, Dept. of Biochemistry, Biozentrum, Medicine, National Cheng-Kung University, University of Basel, Switzerland Tainan, Taiwan HELENIUS, ARI, Dept. of Cell Biology, Yale Univer- JtN, ALBERT, Physical Sciences Laboratory, DCRT, sity, New Haven, Connecticut National Institutes of Health, Bethesda, Maryland HENIS, YOAV I., Dept. of Biochemistry, Faculty of Life JIUING, TAMAS, Dept. of Physiology and Biophysics, Sciences, Tel Aviv University, Tel Aviv, Israel University of Alabama, Birmingham HERMAN, GARY, Dept. of Biochemistry and JOHNSON, ARTHUR, Texas A&M University, Health Biophysics, Hormone Research Institute, Univer- Science Center, College Station sity of California, San Francisco JOYCE, PAUL, Dept. of Chemistry and Biochemistry, HERSKOWITZ, IRA, Dept. of Biochemistry, University Concordia University, Montreal, Canada of California, San Francisco KAISER, CHRIS, Dept. of Biology, Massachusetts In- HIBBS, ALAN, Brisbane Hospital, Queensland Institute stitute of Technology, Cambridge of Medical Research, Herston, Brisbane, Australia KALIES, LAI-UWE, Dept. of Cell Biology, Harvard Ho, SIEW, Graduate Program in Immunology, Harvard Medical School, Boston, Massachusetts Medical School, Massachussetts General Hospital KANDROR, KONSTANTIN, Dept. of Biochemistry, Bos- Cancer Center, Charlestown, Massachusetts ton University Medical School, Boston, Massachu- HOCHSTRASSER, MARK, Dept. of Biochemistry and setts Molecular Biology, University of Chicago, Illinois AH KANG, HYUN, Protein Engineering Research HOLLOWAY, BRIAN, Zeneca Pharmaceuticals, Mac- Group, Genetic Engineering Research Institute, clesfield, Cheshire, United Kingdom Taejon, Korea nORWICH, ARTHUR, Dept. of Molecular Genetics, KANNER, ELLIOT,Dept. of Cellular Biophysics, Rock- Howard Hughes Medical Institute, Yale Univer- efeller University, New York, New York sity School of Medicine, New Haven, Connecticut KAPLAN, ARNOLD, Dept. of Biological Sciences, Uni- HOWELL, KATHRYN, Dept. of Cellular and Structural versity of Arkansas, Fayetteville Biology, University of Colorado School of Medi- KARLSSON, LARS, Dept. of Immunology, R.W. cine, Denver Johnson Pharmaceutical Research Institute, HUHSE, BETTINA, Zellbiochemie, Ruhr-Universitat Scripps Research Institute, La Jolla, California Bochum, Germany KASSESSINOFF,TATIANA, Laboratory of Molecular Im- HURT, EDUARD, European Molecular Biology munology, National Heart, Lung, and Blood In- Laboratory, Heidelberg, Germany stitute, National Institutes of Health, Bethesda, HURTLEY, STELLA, Science, Cambridge, United King- Maryland dom KEEGSTRA, KENNETH, MSU-DOE Plant Research HUTrNER, WIELAND, Institute for Neurobiology, Uni- Laboratory, Michigan State University, East Lan- versity of Heidelberg, Germany sing IMAMOTO, NAOKO, Dept. of Molecular and Cellular KELLY, REGIS, Dept. of Biochemistry and Biophysics, Biology, Osaka University, Suita Osaka, Japan University of California, San Francisco ITO, AKIO, Dept. of Chemistry, Kyushu University, KIESEWALTER, FRANK, Instit0t fiir Molekularbiologie Fukuoka, Japan und Biochemie, Freie Universit~it Berlin, Germany SYMPOSIUM PARTICIPANTS ix

KIRCHHAUSEN, TOM, Dept. of Cell Biology, Harvard LISANTI, MICHAEL, Whitehead Institute for Biomedi- Medical School, Boston, Massachusetts cal Research, Cambridge, Massachusetts KNOLL, BRIAN, Dept. of Medicine and Cell Biology, LIu, JUN, Dept. of Pathology, Beth Israel Hospital, Baylor College of Medicine, Houston, Texas Harvard Medical School, Boston, Massachusetts KOLLER, ERICH, La Jolla Cancer Research Founda- LIU, SHu-HuI, Dept. of Microbiology and Immunology tion, California & Pharmacy, University of Cmifornia, San KREIBICH, GERT, Dept. of Cell Biology, New York Francisco University Medical Center, New York LODISH, HARVEY,Dept. of Biology, Massachusetts In- KUNAU, WOLF, Zellbiochemie, Ruhr-Universitat stitute of Technology, Cambridge Bochum, Germany LOEFFLER, KATI, Dept. of Pharmacology and Toxicol- KUNZLER, MARKUS,Dept. of Molecular and Cell Biol- ogy, Michigan State University, East Lansing ogy, University of California, Berkeley LORD, MIKE, Dept. of Biological Sciences, University KURODA, YOICHIRO, Dept. of Neurochemistry, Tokyo of Warwick, Coventry, United Kingdom Metropolitan Institute for Neuroseience, Japan LOUVARD, DANIEL, Dept. of Biology, Institut Pasteur, KUTAY, ULRtKE,Dept. of Cell Biology, Harvard Medi- Paris, France cal School, Boston, Massachusetts LOWE, JAN, Max-Planck Institute for Biochemistry, LAFER, EILEEN, Dept. of Biotechnology, University of Abteilung Strukturforschung, Martinsried, Texas Health Sciences Center, San Antonio Germany LAFONT, FRANK, Cell Biology Programme, European LUDT, JANNIKKE, Dept. of Biochemistry, Norwegian Molecular Biology Laboratory, Heidelberg, Hospital, Institute for Cancer Research, Germany Montebelio, Oslo, Norway LAHTINEN, ULLA, Ludwig Institute for Cancer Re- MACHOLD, ROBERT, Dept. of Biology, Massachusetts search, Sweden Institute of Technology, Cambridge LAMAZE, CHRISTOPHE, Dept. of Cell Biology, The MAJDALANI, NADIM, Dept. of Medical Microbiology Scripps Research Institute, La Jolla, California and Immunology, Texas A&M University, College LAMPERT, ROBERT, Dept. of Medicine, Baylor College Station of Medicine, Houston, Texas MALETIC-SAVATIC, MIRJANA, Cold Spring Harbor LANZREIN, MARKUS,Dept. of Biochemistry, The Nor- Laboratory, Cold Spring Harbor, New York wegian Radium Hospital, Montebello, Oslo MALHOTRA, VIVEK, Dept. of Biology, University of LAURING, BRETT, Dept. of Cell Biology, New York California at San Diego, La Jolla University Medical Center, New York MARKS, MICHAEL, Cell Biology and Metabolism LAWRENCE, SUSAN,Plant Science Center, Cornell Uni- Branch, National Institute of Child Health and versity, Ithaca, New York Human Development, National Institutes of LE BIVlC, ANDRE, Genetique et Physiologie du Devel- Health, Bethesda, Maryland oppement, Parc Scientifique de Luminy, Univer- MARTIN, THOMAS, Dept. of Zoology Research, Uni- site D-Aix-Marseille, Marseille, France versity of Wisconsin, Madison LEBEAU, MARIE-CLAIRE, INSERM, Le Kremlin MATTA/, IAIN, Gene Expression Programme, Euro- Bicetre, France pean Molecular Biology Laboratory, Heidelberg, LEE, DO, Dept. of Cell Biology, Harvard Medical Germany School, Boston, Massachusetts MAqq'EI, DENISE, Dept. of Immunology/Experimental LEE, KEE NYUNG, Protein Engineering Group, Genet- Parasitology, Institut Pasteur, Paris, France ic Engineering Research Institute, Taejon, Korea MATTHEWS, PAUL, Montreal Neurological Institute, LEFEBVRE, YVONNE, Dept. of Medicine and McGill University, Quebec, Canada Biochemistry, University of Ottawa, Ottawa Civic MCCARTHY, J. BRIAN, Dept. of Neurobiology, State Hospital, Ontario, Canada University of New York, Stony Brook LEWIS, MICHAEL, Division of Cell Biology, MRC MCCLINTOCK, TIM, Dept. of Physiology, University of Laboratory of Molecular Biology, Cambridge, Kentucky College of Medicine, Lexington United Kingdom MCGRAW, PATRICIA, Dept. of Biological Sciences, LIAO, SHUREN,Texas A&M University Health Science University of Maryland, Baltimore Center, College of Medicine, College Station MCNALLY, TERESA, Abbott Laboratories, Abbott LIN, YAO, Dept. of Microbiology and Immunology, Park, Illinois Vanderbilt University School of Medicine, Nash- MEHTA, PRASHANT, Dept. of Neuropharmacology, ville, Tennessee Scripps Research Institute, La Jolla, California LINDQUIST, SUSAN, Dept. of Molecular Genetics and MELLMAN, IRA, Dept. of Cell Biology, Yale University Cell Biology, University of Chicago, Illinois School of Medicine, New Haven, Connecticut x SYMPOSIUM PARTICIPANTS

MICHAELIS, SUSAN, Dept. of Cell Biology and NEWITI', JOHN, NIDDK/GBB, National Institutes of Anatomy, Johns Hopkins School of Medicine, Bal- Health, Bethesda, Maryland timore, Maryland NEWMAN, LORI, Lab. of Molecular Neuro-Oncology, MICHAUD, WILLIAM, Dept. of Biological Sciences, Rockefeller University, New York, New York University of Maryland, Baltimore NIELSEN, ERIK, MSU-DOE Plant Research Labo- MIHARA, KAYSUYOSHI, Dept. of Molecular Biology, ratory, Michigan State University, East Lansing Graduate School of Medical Science, Kyushu Uni- NOSAKA, MICHIKO, Dept. of Material and Biological versity, Maidashi, Higashi-ku, Fukuoka, Japan Engineering, Sasebo National College of Technol- MILISAV-RIBARIC, IRINA, Dept. of Pathology, Univer- ogy, Sasebo, Nagasaki, Japan sity of Cambridge, United Kingdom NOSSAL, RALPH, Dept. of Physical Sciences/Computer MINSKOFF, STACEY, Dept. of Cell Biology, Yale Uni- Research and Technology, National Institutes of versity School of Medicine, New Haven, Con- Health, Bethesda, Maryland necticut NOVtCK, PETER, Dept. of Cell Biology, Yale University MISRA, RAJEEV, Dept. of Microbiology, Arizona State School of Medicine, New Haven, Connecticut University, Tempe NUTLEY, WILLIAM, Dept. of Biology, University of MOORE, MARY, Dept. of Cell Biology, Baylor College California at San Diego, La JoUa of Medicine, Houston, Texas O'BRIEN, LUCY, Dept. of Physiology, University of MOORE, ROBERT, Dept. of Cancer Research, Eli Lilly California, San Francisco and Company, Indianapolis, Indiana O'HALLORAN, TERRY, Dept. of Cell Biology, Duke MORRIS, N. RONALD, Dept. of Pharmacology, Univer- University Medical Center, Durham, North Caro- sity of Medicine and Dentistry of New Jersey, lina Robert Wood Johnson Medical School, Piscataway OHSUMI, YOSHINORI, Dept. of Biology, University of MOSTOV, KEITH, Dept. of Anatomy, University of Cal- Tokyo, College of Arts & Sciences, Komaba, ifornia, San Francisco Meguro-ku, Japan MUHLBERG, AMY, Dept. of Cell Biology, The Scripps OKAMOTO, PATRICIA, Cell Biology Group, Worcester Research Institute, La Jolla, California Foundation for Experimental Biology, Shrews- MUNRO, SEAN, Dept. of Cell Biology, MCR bury, Massachusetts Laboratory of Molecular Biology, Cambridge, OLIVER, JASON, University of Manchester, School of United Kingdom Biological Sciences, Manchester, United Kingdom MUTH, THEODORE, Dept. of Cell Biology, Yale Uni- OLKKONEN, VESA, Dept. of Biochemistry, National versity, New Haven, Connecticut Public Health Institute, Helsinki, Finland NAKANO, AKIHIKO, Dept. of Plant Sciences, Univer- OMURA, TSUNEO,Dept. of Molecular Biology, Kyushu sity of Tokyo, Japan University, Higashi-ku, Fukuoka, Japan NAPIER, JOHNATHAN, Dept. of Cell Biology, Univer- OUWENDIJK, JOHANNA, Dept. of Cell Biology and sity of Bristol, United Kingdom Histology, University of Nijmegen, The Nether- NAYLOR, DEAN, Dept. of Horticulture, Viticulture and lands Oenology, University of Adelaide, Glen Asmond, PADH, HARISH,Dept. of Biotechnology, Northwestern Australia University, Evanston, Illinois NEALE, ELAINE, Dept. of Developmental Neurobiol- PAEK, INBOK, Dept. of Cellular Biochemistry and ogy, National Institute of Child Health and Hu- Biophysics, Memorial Sloan-Kettering Cancer man Development, National Institutes of Health, Center, New York, New York Bethesda, Maryland PALADE, GEORGE, Dept. of Medicine, University of NEDERLOF, PETRA, Dept. of Structural Biology, Max- California at San Diego, La Jolla Planck Institute for Biochemistry, Martinsried, PANTE, NELLY, Biozentrum, M.E. Muller Institute, Germany University of Basel, Switzerland NEHRBASS, ULF, Dept. of Cell Biology, Rockefeller PELHAM, HUGH, MRC Laboratory of Molecular Biol- University, Howard Hughes Medical Institute, ogy, Cambridge, United Kingdom New York, New York PETANCESKA, SUZANA, Dept. of Molecular and Cel- NEIMAN, AARON, Dept. of Biochemistry, State Univer- lular Neuroscience, Rockefeller University, New sity of New York, Stony Brook York, New York NELSON, W. JAMES, Dept. of Molecular and Cellular PETERS, PETER, Dept. of Cell Biology, Medical School, Physiology, Stanford University, Stanford, Califor- Utrecht University, The Netherlands nia PETERSON, PER, R.W. Johnson Pharmaceutical Re- NEUPERT, WALTER, Physiologische Chemic, Univer- search Institute, San Diego, California sity of Munich, Germany PETROU, CONSTANTIA, Dept. of Molecular and Cel- SYMPOSIUM PARTICIPANTS xi

lular Toxicology, Harvard University School of ROSA, JOSE LUXS, Dept. of Molecular Oncology, Public Health, Boston, Massachusetts Bristol-Myers Squibb Pharmaceuticals, Princeton, PETrERSSON, RALF, Ludwig Institute for Cancer Re- New Jersey search, Karolinska Institute, Stockholm, Sweden ROTHMAN, JAMES, Dept. of Cellular Chemistry and PFEFFER, SUSANNE, Dept. of Biochemistry, Stanford Biophysics, Memorial Sloan-Kettering Cancer University School of Medicine, Stanford, Califor- Center, New York nia ROTTIER, PETER, Dept. of Infectious Diseases and Im- PIERCE, SUSAN, Dept. of Biochemistry, Molecular munology, Utrecht University, Veterinary Faculty, Biology and Cell Biology, Northwestern Univer- The Netherlands sity, Evanston, Illinois ROUSH, DENISE, Dept. of Cellular and Molecular PIETERS, JEAN, Dept. of Immunology, Netherlands Physiology, Yale University, New Haven, Con- Cancer Institute, Amsterdam,The Netherlands necticut PLATH, KATHRIN, Dept. of Stress Proteins, Max- ROWE, TONY, Dept. of Cell Biology, The Scripps Re- Delbriick Center of Molecular Medicine, Berlin, search Institute, La Jolla, California Germany Russ, GUSTAV, Dept. of Cell Biology, National In- PLESS, DOROTHY,Fort Detrick, Frederick, Maryland stitute of Allergy and Infectious Diseases, National PLOEGH, HIDDE, Massachusetts Institute of Technol- Institutes of Health, Bethesda, Maryland ogy, Center for Cancer Research, Cambridge SABATINI, DAVID, Dept. of Cell Biology, New York POGLIANO, JOE, Dept. of Microbiology, Harvard University Medical Center, New York Medical School, Boston, Massachusetts SAITOH, HISATO, Lab. of Molecular Embryology, Na- POHLSCHRODER, MECHTHILD,Dept. of Microbiology tional Institute of Child Health and Human Devel- and Molecular Genetics, Harvard Medical School, opment, National Institutes of Health, Bethesda, Boston, Massachusetts Maryland POD, Mu-MING, Dept. of Biological Sciences, Colum- SAKAGUCHI, MASAO, Dept. of Molecular Biology, bia University, New York, New York Kyushu University, Graduate School of Medical PRINGLE, JOHN, Dept. of Biology, University of North Science, Fukuoko, Japan Carolina, Chapel Hill SALOHEIMO, MARKKU,VTT Biotechnology and Food RAPOPORT, THOMAS, Dept. of Cell Biology, Harvard Research, Espoo, Finland Medical School, Boston, Massachusetts SANTINI, FRANCESCA,Dept. of Pharmacology, Jeffer- RAYNER, JULIAN, Dept. of Cell Biology, MRC son Cancer Institute, Thomas Jefferson University, Laboratory of Molecular Biology, Cambridge, Philadelphia, Pennsylvania United Kingdom SAUCAN, LUCIAN, Dept. of Medicine, University of REDDY, PADMALATHA,Dept. of Microbiology, Boston California at San Diego, La Jolla University School of Medicine, Boston, Massachu- SCHATZ, GOTTFRIED, Basel Abteilung Biochemistry, setts Biozentrum University, Basel, Switzerland REINSTEIN, JOCHEN, Max-Planck Institute for Im- SCHEKMAN, RANDY, Dept. of Biochemistry and munobiology, Abteilung Physikalische Biochemie, Molecular Biology, University of California, Dortmund, Germany Berkeley REN, MINDONG,Dept. of Cell Biology, New York Uni- SCHELLER, RICHARD,Dept. of Molecular and Cellular versity Medical Center, New York Physiology, Howard Hughes Medical Institute, RENFREW-HAFT, CAROL, Diabetes Branch, NIDDK, Stanford University, Stanford, California National Institutes of Health, Bethesda, Maryland SCHERER, PHILIPP, Dept. of Research, Whitehead In- REXACH, MICHAEL,Dept. of Cell Biology, Rockefeller stitute for Biomedical Sciences, Cambridge, Mass- University, New York, New York achusetts RIKIHISA, YASUKO, Dept. of Veterinary Pathobiology, SCHMID, SANDRA, The Scripps Research Institute, La Ohio State University, Columbus Jolla, California RINDLER, MICHAEL,Dept. of Cell Biology, New York SCHNELL, DANIEL, Dept. of Biological Sciences, Rut- University Medical Center, New York gers University, Newark, New Jersey RIZZOLO, LAWRENCE,Dept. of Surgery, Yale Univer- SCHNITZER, JAN, Dept. of Pathology, Harvard Medical sity School of Medicine, New Haven, Connecticut School, Beth Israel Hospital, Boston, Massachu- ROBERTSON, MIRANDA, Garland, London, United setts Kingdom SCHOREY, CRISLYN,Dept. of Cell Biology, Washington ROHRER, JACK, Dept. of Hematology/Oncology, University School of Medicine, St. Louis, Missouri Washington University School of Medicine, St. SCHWARZ, THOMAS, Dept. of Molecular and Cellular Louis, Missouri Physiology, Stanford University Medical Center, xii SYMPOSIUM PARTICIPANTS

Stanford, California SODHOF, THOMAS, Dept. of Molecular Genetics, Uni- SHEETZ, MICHAEL, Dept. of Cell Biology, Div. of versity of Texas Southwestern Medical Center, Physiology, Duke University Medical Center, Dur- Dallas ham, North Carolina SUSAN, PETER, Dept. of Anatomy and Cell Biology, SHEN, BETTY, Dept. of Molecular Biology and Micro- University of Florida, Gainesville biology, Yale University, New Haven, Connecticut SYIN, CHIANG, Dept, of Parasitic Biology and SHIA, MICHAEL,Dept. of Medicine, Boston University Biochemistry, Food and Drug Administration, School of Medicine, Massachusetts RockviUe, Maryland SHRAIMAN, BORIS, Theoretical Physics Section, AT&T SZlLARD, RACHEL, Dept. of Anatomy and Cell Biol- Bell Laboratories, Murray Hill, New Jersey ogy, University of Alberta, Edmonton, Canada SHVNG, SHOW-LING, Dept. of Cell Biology and SZTUL, ELIZABETH, Dept. of Molecular Biology, Physiology, Washington University School of Princeton University, Princeton, New Jersey Medicine, St. Louis, Missouri TABAK, HENK, Dept. of Biochemistry, E.C. Slater In- SIGGIA, ERIC, Dept. of Physics, Cornell University, stitute, Amsterdam, The Netherlands Ithaca, New York TAHARA, MASAHIRO, Dept. of Theoretical and Physi- SIJTS, ALICE,Dept. of Internal Medicine, Yale Univer- cal Biology, National Institute of Child Health and sity School of Medicine, New Haven, Connecticut Human Development, National Institutes of SIMON, JEAN-PIERRE, Dept. of Cell Biology, New York Health, Bethesda, Maryland University Medical Center, New York TAKANO, EMIKO, Institute for Virus Research, Kyoto SIMON, SANFORD, Dept. of Cellular Biophysics, Rock- University, Kyoto, Sakyo-ku, Japan efeller University, New York, New York TAMPE, ROBERT, Max-Planck Institute for Biochem- SIMON, SIDNEY, Dept. of Neurobiology, Duke Univer- istry, Martinsried, Germany sity Medical Center, Durham, North Carolina TATU, UTPAL, Dept. of Cell Biology, Yale University SIMONS, KAI, Cell Biology Program, European School of Medicine, New Haven, Connecticut Molecular Biology Laboratory, Heidelberg, TERBUSH, DANIEL, Dept. of Cell Biology, Yale Uni- Germany versity School of Medicine, New Haven, Con- SKACH, WILLIAM, Dept. of Molecular and Cellular necticut Engineering, University of Pennsylvania, Philadel- THOMPSON, NICOLA, Dept. of Cellular Science, Glaxo phia Research and Development, Greenford, Middle- SKEHEL, JOHN, MRC National Institute for Medical sex, United Kingdom Research, London, United Kingdom TING, TING, Dept. of Molecular and Cellular Physiol- SLEPNEV, VLADIMIR,Dept. of Cell Biology, Yale Uni- ogy, Stanford University Medical Center, Stan- versity School of Medicine, New Haven, Con- ford, California necticut TOKATLIDIS, KOSTAS, Dept. of Biochemistry, SMITH, CORINNE, Dept. of Biochemistry, School of Biozentrum, University of Basel, Switzerland Medical Sciences, University of Bristol, United TORGERSON, TROY, Dept. of Microbiology and Im- Kingdom munology, Vanderbilt University School of Medi- SONG, WENXIA, Dept. of Biochemistry, Molecular cine, Nashville, Tennessee Biology, and Cell Biology, Northwestern Univer- TRANEL, PATRICK, Plant Research Laboratory, sity, Evanston, Illinois Michigan State University, East Lansing SPRINGER, SEBASTIAN, Institute of Molecular Medi- TROWBRIDGE, IAN, Dept. of Cancer Biology, Salk In- cine, John Radcliffe Hospital, University of Ox- stitute, La Jolla, California ford, United Kingdom TUVIN, MICHAEL,Dept. of Medicine, Baylor College SPUDICH, JAMES, Dept. of Biochemistry, Stanford Uni- of Medicine, Houston, Texas versity School of Medicine, Stanford, California ULRICH, HELLE, Dept. of Chemistry, University of STILLMAN, BRUCE, Cold Spring Harbor Laboratory, California, Berkeley Cold Spring Harbor, New York URBAN, JOERG, Max-Delbr0ck Center for Molecular STIRLING, COLIN, Dept. of Biochemistry and Medicine, Berlin, Germany Molecular Biology, University of Manchester, VALLEE, RICHARD, Dept. of Cell Biology, Worcester United Kingdom Foundation for Experimental Biology, Shrews- STRAMBIO DE CASTILLIA, CATERINA, Dept. of Cell bury, Massachusetts Biology, Rockefeller University, New York, New VARSHAVSKY, ALEXANDER,Dept. of Biology, Califor- York nia Institute of Technology, Pasadena SUBRAMANI, SURESH, Dept. of Biology, University of VERHEY, KRISTEN, Dept. of Cell Biology, Harvard California at San Diego, La Jolla Medical School, Boston, Massachusetts SYMPOSIUM PARTICIPANTS xiii

VILLADANGOS,JosE, Dept. of Cancer Research, Mass- WILKINSON, BARRIE, School of Biological Sciences, achusetts Institute of Technology, Cambridge University of Manchester, United Kingdom WALTER, PETER, Dept. of Biochemistry and WILSON, RICHARD, Dept. of Biochemistry, School of Biophysics, University of California, San Francisco Biological Sciences, University of Manchester, WARD, COLIN, Dept. of Biomolecular Engineering, United Kingdom CSIRO, Parkville, Victoria, Australia WINCKLER, BETTINA, Dept. of Biological Sciences, WARREN, GRAHAM, Dept. of Cell Biology, Imperial Columbia University, New York, New York Cancer Research Fund, London, United Kingdom WOZNIAK, MAGDALENA, Dept. of Pharmacology, WARREN, ROBIN, Dept. of Cell Biology, Oregon Vanderbilt University Medical School, Nashville, Health Sciences University, Portland Tennessee WEIS, KARSTEN, Gene Expression Programme, Euro- WOZNIAK, RICHARD,Dept. of Anatomy and Cell Biol- pean Molecular Biology Laboratory, Heidelberg, ogy, University of Alberta, Edmonton, Canada Germany Xu, HUAXI, Dept. of Molecular and Cellular Neuro- WELLNER, ROBERT, Dept. of Cell Biology and science, Rockefeller University, New York, New Biotechnology, U.S. Army Medical Research In- York stitute of Infectious Diseases, Fort Detrick, YAVER, DEBBIE, Dept. of Molecular Biology, Novo Frederick, Maryland Nordisk Biotech, Inc., Davis, California WENDLAND, BEVERLY, Dept. of Cellular and YEAMAN, CHARLES, Dept. of Cell Biology and Molecular Medicine, Howard Hughes Medical In- Anatomy, Cornell University Medical College, stitute, University of California at San Diego, La New York, New York Jolla YOKOYAMA, NOBUHIKO, Dept. of Molecular Biology, WESSEL, GARY, Dept. of Molecular, Cellular Biology Kyushu University, Fukuoka, Japan and Biochemistry, Brown University, Providence, YORK, IAN, Dept. of Lymphocyte Biology, Dana- Rhode Island Farber Cancer Institute, Boston, Massachusetts WESSLING-RESNICK, MARIANNE, Dept. of Nutrition, YOUNG, JAMES, Dept. of Biochemistry, McMaster Uni- Harvard School of Public Health, Boston, Massa- versity, Hamilton, Ontario, Canada chusetts ZENG, JIANBO, Dept. of Cell Biology, New York Uni- WHITE, JUDITH, Dept. of Cell Biology, University of versity Medical Center, New York Virginia Health Sciences Center, Charlottesville ZERIAL, MARINO, Eurpopean Molecular Biology WHITTERS, ERIC, Molecular Biology Institute, Univer- Laboratory, Heidelberg, Germany sity of Oregon, Eugene ZHANG, HONG, Dept. of Biochemistry, University of WICKNER, WILLIAM, Dept. of Biochemistry, Dart- Wisconsin, Madison mouth Medical School, Hanover, New Hampshire ZIMMERBERG, JOSHUA, Dept. of Theoretical and WIEDMAN, MARTIN, Memorial Sloan-Kettering Cancer Physical Biology, National Institutes of Health, Center, New York, New York Bethesda, Maryland First row: G. Blobel, M. Sheetz, G. Kreibich; W. Almers, J. Rothman Second row: K. Simons; M.-J. Gething, B. Stillman; P. Peterson, R. Kelly Third row: T. Rapoport, M. Brunner, W. Neupert; K. Howell, T. Kirchhausen Fourth row: After Dorcas Cummings lecture; Wine and cheese First row: J, White; L. Rizzolo; R. Pettersson, L. Gerace Second row: 1. Trowbridge; S. Gottesman; H. Pelham; M. Hatanaka Third row: P. Walter; P. deCamilli, M. Farquhar; W. Wickner, A. Goldberg Fourth row: Coffee break, Grace; I. Herskowitz First row: D. Louvard, R. Kelly, M.-J. Gething, R. Anderson, K. Simons; R. Pettersson, B. Stillman Second row: Y. Ohsumi, A. Nakano, R. Schekman; K. Mostov, M. Zerial, H. Lodish Third row: S. Block entertains; V. Malhotra, M. Brunner Fourth row: Picnic, Blackford lawn First row: M. Moore, T. O'Halloran; D. TerBush; G. Schatz Second row: J. Donaldson; T. Sfidhof; R. Schekman, G. Palade Third row: R. Scheller; K. Cline, R. Martienssen; G. Dreyfuss Fourth row: A. Johnson; A. Helenius, I. Mellman; J. Beckwith Foreword

The way individual cells look and function follows directly from the program of cell-type- dependent gene expression and the nature of the proteins that are expressed. Once synthesized, however, the proteins themselves take on a life of their own that is determined by their amino acid sequence and their environment. Proteins have varied lifetimes, cellular locations, and interactions with other proteins or cellular structures such as membranes, each of which can influence cell physiology directly. Thus, the life of proteins in the cell is an important topic that has deserved much attention. At the last Symposium that dealt with these issues, in 1981, the focus was on the structure of the cytoskeleton and general observations of protein trafficking within the cell, such as cell-surface polarity and axonal transport. The science was dominated by microscopy and was primarily observational. As pointed out by Jim Watson in his Foreword to that Symposium volume, the field was crying out for an in- flux of biochemistry and genetics to present a more detailed understanding of the mechanisms of protein stability, movement, and organization. With remarkable results, the biochemists and geneticists heeded the call. During the last 14 years, we have witnessed a dramatic increase in knowledge about the mechanisms of protein movement within the cell. It was therefore prime time to dedicate the Symposium to the dynamic aspects of protein function. The title "protein kinesis" was invented to reflect the many aspects of protein modification, trafficking, stability, localization, and organization. Knowledge of the mechanisms of protein transport either through the endoplasmic reticulum and Golgi to the cell surface, or into the nucleus, has progressed rapidly in recent years. Our grasp of the subject is sufficient that we can begin to appreciate links between these processes in different cell types and to understand how protein kinesis influences human disease processes, a topic that I suspect will dominate future discussion and directions. I am very indebted to Gtinter Blobel, Jim Rothman, Randy Schekman, Mary-Jane Ge- thing, Susan Gottesman, Ulrich Hartl, Ira Mellman, and Ron Laskey for advice and guid- ance on the areas covered in this Symposium. The formal program consisted of 87 talks and 189 poster presentations, and the meeting attracted a total of 389 participants. Outstanding first-night introductory lectures by Giinter Blobel, Jim Rothman, Per Peterson, and Jim Spudich set an exceptionally high standard that was maintained throughout the meeting. Furthermore, Giinter Blobel delivered a masterful Dorcas Cummings lecture for the benefit of members of the local community, and George Palade provided a thoughtful and insightful summary of the entire meeting. This year also saw the inaugural Reginald G. Harris Lecture, given by Randy Schekman. When he was Director of the Biological Laboratories at Cold Spring Harbor, Reginald Harris started the Symposia in 1933 to foster a quantitative approach to biology and to promote interactions between scientists from diverse fields. The modern-day success of these meetings stems directly from his foresight. Indeed, many Symposia have profoundly influenced the future of biology in general. Essential funds that supported this meeting were obtained from the U.S. Department of Energy and the National Institutes of Health. Additional support came from our ever more important and needed Corporate Sponsors: American Cyanamid Company; Amgen Inc.; BASF Bioresearch Corporation; Beckman Instruments, Inc.; Becton Dickinson and Compa- ny; Bristol-Myers Squibb Company; Chugai Pharmaceutical Co., Ltd.; Chugai Research In- stitute for Molecular Medicine, Inc.; Diagnostic Products Corporation; The Du Pont Merck Pharmaceutical Company; Forest Laboratories, Inc.; Genentech, Inc.; Glaxo; Hoffmann-La Roche Inc.; Human Genome Sciences, Inc.; Johnson & Johnson; Kyowa Hakko Kogyo Co., Ltd.; Life Technologies, Inc.; Marion Merrell Dow Inc.; Mitsubishi Kasei Institute of Life Sciences; Monsanto Company; New England BioLabs, Inc.; Oncogene Science, Inc.; Pall Corporation; The Perkin-Elmer Corporation; Pfizer Inc.; Research Genetics, Inc.; Sandoz Research Institute; Schering-Plough Corporation; SmithKline Beecham Pharmaceuticals; Sumitomo Pharmaceuticals Co., Ltd.; The Upjohn Company; The Wellcome Research Laboratories, Burroughs Wellcome Co.; Wyeth-Ayerst Research; and Zeneca Group PLC. Once again, I thank the efficient staff in our meetings and courses office, under the very capable direction of David Stewart, who now assumes responsibility for directing our large

xix FOREWORD meetings and courses program that emerged from a single annual Symposium begun 63 years ago. In particular, Diane Tighe efficiently managed the many logistical aspects of meetings registration; Mary Horton ably handled the various grant applications; and Lee Martin did an excellent job of preparing the abstract book. As usual, Herb Parsons and his staff provided excellent audiovisual assistance. The organization of this meeting greatly relied on the marvelous work of my assistant Delia King. Finally, it was again a pleasure to work with the efficient Laboratory Press, under the direction of John Inglis: particularly Nancy Ford, Patricia Barker, Joan Ebert, Elaine Gaveglia, and Susan Schaefer. Their efficiency made this volume possible.

Bruce Stillman March 1996 Contents

Symposium Participants V Foreword xix

Protein Translocation in the Endoplasmic Reticulum and Golgi

Unidirectional and Bidirectional Protein Traffic across Membranes G. Blobel 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 Mem- branes 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 Trans- duction H.F. Lodish, D.J. Hilton, U. Klingmiiller, S.S. Watowich, and H. Wu 93 SNAREs Involved in Traffic through the Golgi Complex H.R.B Pelham, D.K. Ban- field, 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-Kri~ger, 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 Receptor- mediated 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, LE. lvanov, M. Ren, J. Zeng, B. Shopsin, D. Hersh, P. Tempst, H. Erdjument-Bromage, xxi ~ii CONTENTS

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 andJ. 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. Ranca~o, 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 Plas- malemmal 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. Friih, 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. PohlschrOder, 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. Siidhof 309 Biogenesis of Neurosecretory Vesicles W.B. Huttner, M. Ohashi, R.H. Kehlenbach, F.A. Barr, R. Bauerfeind, O. Briiunling, D. Corbeil, M. Hannah, H.A. Pasolli, A. Schmidt, A.A. Schmidt, C. Thiele, Y. Wang, A. KrOmer, and H.-H. Gerdes 315 Botulinum and Tetanus Neurotoxins: Emerging Tools for the Study of Membrane Fu- sion R. Jahn, P.L 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. Bon- zelius, 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 CONTENTS xxiii

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. BIond-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 Hspl04 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, L 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. Seemi~ller, 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. L6we 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, L Plonsky, A. Sokoloff, A. Chanturia, and L.V. Chernomordik 589 Role of Water in Short-range Repulsive and Attractive Interactions between Macro- molecules and Bilayers S.A. Simon and T.J. Mclntosh 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 Mem- branes M. Brunner, H.-C. Schneider, R. Lill, and W. Neupert 619 Molecular Dissection of the Mechanism of Protein Import into Chloroplasts F. Tian, Y. ~iv CONTENTS

Ma, A. Kouranov, S.E. LaSala, and D.J. Schnell 629 Characterization of the Yeast and Human Receptors for the Carboxy-terminal Tripep- tide 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, }1. 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. Pihol-Roma, S. Nakielny, Q. Liu, and G. Dreyfuss 663 Nuclear Localization of a Cap-binding Protein Complex E. Izaurralde, C. McGuigan, and L 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. GOrlich 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 L 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 L 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. lkonen 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 Im- munoglobulin 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. Pat- terson, 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. Os- mani, C.H. Roghi, D.A. Willins, S. Beckwith, (7. Goldman, Y. Chiu, M. Xin, B. Liu, and N.R. Morris 813 Summary: G.E. Palade 821

Author Index 833

Subject Index 835