The Reoviridae THE VIRUSES

Series Editors HEINZ FRAENKEL-CONRAT, University of California Berkeley, California ROBERT R. WAGNER, University of Virginia School of Medicine Charlottesville, Virginia

THE HERPESVIRUSES, Volumes I, 2, 3, and 4 Edited by Bernard Roizman

THE REOVIRIDAE Edited by Wolfgang K. Joklik

THE PARVOVIRUSES Edited by Kenneth I. Berns The Reoviridae

Edited by WOLFGANG K. JOKLIK Duke University Medical Center Durham, North Carolina

Springer Science+Business Media, LLC Library of Congress Cataloging in Publication Data Main entry under title: The Reoviridae. (Viruses) Includes bibliographical references and index. 1. Reoviruses. L Joklik, Wolfgang K. II. Series. QR414.R46 1983 576/.64 83-6276 ISBN 978-1-4899-0582-6

ISBN 978-1-4899-0582-6 ISBN 978-1-4899-0580-2 (eBook) DOI 10.1007/978-1-4899-0580-2

© Springer Science+Business Media New York 1983 Originally published by Plenum Press, New York in 1983 Softcover reprint of the hardcover 1st edition 1983

All rights reserved No part of this book may be reproduced, stored in a retrieval system, or transmitted in any form or by any means, electronic, mechanical, photocopying, microfilming, recording, or otherwise, without written permission from the Publisher Con tribu tors

Guido 'Boccardo, Istituto di Fitovirologia Applicata del C.N.R., 10135 To• rino, Italy Bernard N. Fields, Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115; and Depart• ment of Medicine, Brigham and Women's Hospital, Boston, Massa• chusetts 02115 R. I. B. Fran'cki, Department of Plant Pathology, Waite Agricultural Re• search Institute, The University of Adelaide, Adelaide 5064, South Australia Barry M. Gorman, Queensland Institute of Medical Research, Bramston Terrace, Brisbane 4006, Australia Ian H. Holmes, Department of Microbiology, University of Melbourne, Parkville, Victoria 3052, Australia Wolfgang K. Joklik, Department of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina 27710 Marilyn Kozak, Department of Biological Sciences, University of Pitts• burgh, Pittsburgh, Pennsylvania 15260 Peter P. C. Mertens, Animal Virus Research Institute, Pirbright, Woking, Surrey GU24 ONF, United Kingdom Stewart Millward, Department of Biochemistry, McGill University, Mon• treal, Quebec, Canada H3G 1Y6 Christopher C. Payne, Glasshouse Crops Research Institute, Littlehamp• ton, West Sussex BN16 3PU, United Kingdom Robert F. Ramig, Department of Virology and Epidemiology, Baylor Col• lege of Medicine, Houston, Texas 77030 Arlene H. Sharpe, Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115 Aaron J. Shatkin, Department of Cell Biology, Roche Institute of Mole• Biology, Nutley, New Jersey 07110 v vi CONTRIBUTORS

Jill Taylor, Queensland Institute of Medical Research, Bramston Terrace, Brisbane 4006, Australia Peter J. Walker, Queensland Institute of Medical Research, Bramston Ter• race, Brisbane 4006, Australia Helmut Zarbl, Department of Biochemistry, McGill University, Mon• treal, Quebec, Canada H3G 1Y6 Preface

It is now just 20 years since Gomatos and his co-workers at the Rocke• feller University showed that the nucleic acid in reovirus particles is double-stranded RNA (dsRNA). This discovery created great excitement, for dsRNA was at that time under intense investigation as the replicative form of viral genomes consisting of single-stranded RNA. An equally interesting and important finding followed soon after: it was found that the reovirus genome consists, not of a single nucleic acid molecule, but of 10 discrete "segments," each with its specific sequence content and each transcribed into its own messenger RNA. It is clear now that these segments are genes. Not surprisingly, the availability of a viral genome consisting of 10 unlinked genes has permitted some unique lines of in• vestigation in molecular biology. Mammalian and avian reoviruses proved to be but the first of several viruses recognized as sharing Similarity in size and morphology and ge• nomes consisting of 10, II, or 12 separate genes. These viruses are dis• tributed throughout living organisms; among the natural hosts of mem• bers of this virus family are vertebrates, Insects, and plants. Members of the Reoviridae family differ widely in the virulence that they exhibit toward their hosts . .For example, the first discovered mam• malian reovirus literally is, as the name signifies, a "respiratory enteric orphan" virus, that is, a virus unassociated with disease. In contrast, the most recently discovered members of this family, the rotaviruses, which were recognized as comprising a separate genus not much more than a decade ago, cause a disease, infantile gastroenteritis, that, while not se• vere in communities where health care is good, is one of the most im• portant, if not the most important, cause of death in infants under the age of 2 years in less developed countries. Studies of this family of viruses from molecular, biological, and medical perspectives complement each other to an unusual degree because of the unique nature of the genomes of these viruses. The ten chapters in this book describe what is currently known concerning the molecular biology of members of this virus family on the

vii viii PREFACE one hand and the pathogenesis that they exhibit toward their ·hosts on the other. About half the chapters are devoted to the mammalian reo• viruses, which are the Reoviridae that have been studied most intensively in molecular and genetic terms; there are chapters that deal with the nature of reovirus particles and of the reovirus genome, its transcription and replication, the reovirus multiplication cycle, and reovirus genetics. There is also a chapter that describes reovirus pathogenesis in terms of the effects that various individual reovirus gene products exert on host cells. Other chapters deal with the orbiviruses, which include some im• portant animal pathogens such as bluetongue virus; the rotaviruses, im• portant human pathogens; the cytopla~mic polyhedrosis viruses of in• sects; and the plant reoviruses, which include some important plant pathogens.

Wolfgang K. Joklik Contents

Chapter 1 The Members of the Family Reoviridae Wolfgang K. Joklik

I. Introduction ...... 1 n. Properties of Members of the Family Reoviridae ...... 2 A. Possession of Double-Stranded RNA ...... 2 B. Structure ...... 2 C. Nature of the Genome ...... 3 D. Lack of Complete Uncoating ...... 4 E. Possession of Enzymes That Transcribe the Double- Stranded RNA into Messenger RNA ...... 4 III. Unclassified Members of the Family Reoviridae ...... 5 A. Drosophila Virus F and Ceratitis Virus I ...... 5 B. Housefly Virus ...... 5 C. Chum Salmon Virus ...... 6 IV. Other Viruses That Contain Double-Stranded RNA ...... 6 References ...... 6

Chapter 2 The Reovirus Particle Wolfgang K. Joklik

I. Introduction ...... 9 n. Morphology ...... 12 A. Reovirus Particles ...... 12 B. Cores ...... 14 C. Top-Component Particles ...... 16 m. Purification ...... 17 IV. Measurement ...... 17 A. Plaque Formation ...... 17 B. Hemagglutination ...... 18

ix x CONTENTS

C. Enumeration of Virus Particles by Measurement of Optical Density...... 18 V. Reovirus Genome ...... 19 VI. Proteins of Reovirus ...... 22 A. Outer Capsid Shell ...... 25 B. Inner Capsid Shell: The Core ...... 28 C. Nonstructural Reovirus Proteins ...... 30 D. Modification of Reovirus Proteins ...... 31 E. Mechanisms of Chemical and Physical Inactivation of Reovirus Particles ...... ;...... 32 VII. Information Content of the Ten Reovirus Genes ...... 33 VIII. Genetic Relatedness among the Three Serotypes of Reovirus ...... 35 IX. Nature of the Antibodies Elicited by Reovirus-Specified Proteins ...... 40 X. Sequence of Reovirus Genes and Messenger RNAs ...... 44 A. Sequence Identity of the Plus Strands of Reovirus Genes and of Their Transcripts ...... 46 B. Sequences at the Termini of the Ten Genes of Reovirus Serotype 3 ...... 47 C. Examination of the 5' -Terminal and 3' -Terminal Sequences for Potentially Functional Features ...... 50 D. Sequences at the Termini of the L3, M3, S1, and S2 Genes of Reovirus Serotypes I, 2, and 3 ...... 54 E. Cloning the Reovirus Genes: The Sequence of the Cloned Gene of Serotype 3 ...... 59 XI. Reovirus Oligonucleotides ...... 60 XII. Conversion of Reovirus Particles to Cores and Activation of the Transcriptase ...... 61 XIII. Enzymes in Reovirus ...... 63 XIV. Reovirus Mutants That Contain Fewer Than Ten Genes ...... 68 References ...... 70

Chapter 3 Biochemical Aspects of Reovirus Transcription and Translation Aaron r Shatkin and Marilyn Kozak

I. Introduction ...... 79 II. Transcription ...... 80 A. Discovery of Reovirus-Associated RNA Polymerase ...... 80 B. Reovirus Messenger RNA Synthesis in Infected Cells ...... 81 C. Properties of the Virion Polymerase ...... 83 D. Polypeptides in Transcribing Cores ...... 85 CONTENTS xi

E. Core Enzymes That Synthesize Messenger RNA 5'- Terminal Caps ...... 87 III. Translation of Reovirus Messenger RNAs ...... 90 A. Protein Synthesis in Virus-Infected Cells ...... 91 B. Translation of Reovirus Messenger RNAs in Vitro ...... 93 IV. Closing Thoughts ...... 99 References ...... ,...... 100

Chapter 4 The Reovirus Multiplication Cycle Helmut Zarbl and Stewart Millward

I. Introduction ...... 107 II. Production of Infectious Virions ...... 108 III. Events in the Infected Cell ...... 109 A. Adsorption ...... 110 B. Penetration and Uncoating ...... 113 C. Transcription of the Parental Genome ...... 121 D. Formation of Progeny Double-Stranded RNA ...... 138 E. Transcription from the Progeny Double-Stranded RNA Genome ...... 155 F. Translation of Reovirus Messenger RNAs ...... 159 G. Effects on Host-Cell Functions ...... 173 H. Morphogenesis and Maturation ...... 176 IV. Summary of the Reovirus Life Cycle ...... 182 References ...... 183 Addendum ...... 195

Chapter 5 Genetics of Reoviruses Robert F. Ramig and Bernard N. Fields

I. Introduction ...... 197 II. Nature of the Reovirus Genome ...... 198 III. Types of Mutants ...... 199 A. Temperature-Sensitive Mutants ...... 199 B. Deletion Mutants ...... 201 C. Other Genetic Markers ...... 201 IV. Genetic Interactions ...... 202 A. Complementation ...... 202 B. Recombination (Reassortment) ...... 203 xii CONTENTS

V. Nongenetic Interactions ...... 206 A. Interference ...... 207 B. Extragenic Suppression of ts Phenotype ...... 210 VI. Physical Map of the Reovirus Genome ...... 216 A. tsA ...... 220 B. tsB ...... 221 C. tsC ...... 222 D. tsD ...... 222 E. tsE ...... 223 F. tsF ...... 223 G. tsG ...... 224 H. tsH ...... 224 I. tsI ...... 224 J. tsJ ...... 225 K. Summary ...... 225 References ...... 225

Chapter 6 Pathogenesis of Reovirus Infection Arlene H. Sharpe and Bernard N. Fields

I. Introduction ...... 229 A. Reovirus Structure ...... 230 B. Reovirus Genetics ...... 230 C. General Approach for Genetic Analysis of Reovirus Virulence ...... 231 II. Overview of the Stages in Reovirus Pathogenesis ...... 233 III. Entry ...... 234 IV. Spread ...... 237 V. Immune Response ...... 238 A. Humoral Immunity ...... 238 B. Cellular Immunity ...... 240 C. Interferon ...... 249 D. Thymosin ...... 250 VI. Virus-Hos.t Cell Interactions ...... 251 A. Tissue Tropism: Virus-Receptor Interactions ...... 251 B. Viral Modification of Host Cells: Alteration of Cellular Metabolism ...... 260 C. Viral Modification of Host Cells: Cytoskeletal Organization ...... 264 VII. Persistent Infection ...... 268 VIII. Molecular Epidemiology of Mammalian Reoviruses ...... 275 IX. Concluding Remarks ...... 276 References ...... 278 CONTENTS xiii

Chapter 7 Orbiviruses Barry M. Gorman, Till Taylor, and Peter f. Walker I. Introduction ...... 287 A. Classification of Orbiviruses ...... 288 B. Disease Relationships ...... 291 C. Isolation of Orbiviruses and the Relationships among Them ...... 292 D. Physicochemical Properties Used in Classification of Orbiviruses ...... 307 II. Structural Components and Their Functions ...... 310 A. Morphology of Orbiviruses ...... 310 B. Orbivirus Genome ...... 314 C. Gene Products ...... 318 D. Comments on the Comparative Structures of Reoviruses and Orbiviruses ...... 323 E. Polypeptides Involved in Immunological Reactions ...... 324 III. Replication ...... 326 A. Cell Culture ...... 326 B. Thin-Section Electron Microscopy ...... 326 C. RNA Synthesis ...... 330 D. Protein Synthesis ...... 331 IV. Genetics ...... 332 A. Temperature-Sensitive Mutants of Orbiviruses ...... 332 B. Gene Reassortment ...... 334 C. Mapping Gene Function ...... 337 V. Genetic Relationships among Orbiviruses ...... 338 A. Comparison of Genome Segments of Orbiviruses ...... 338 B. Comparison of Orbivirus Genomes by RNA-RNA Hybridization ...... 339 C. Oligonucleotide Fingerprinting ...... 341 VI. Speciation in Orbiviruses ...... 344 VII. Conclusion ...... 345 References ...... 346

Chapter 8 Rotaviruses Ian H. Holmes

I. Introduction 359 II. Virions ...... 360 A. Morphology ...... 360 B. Physicochemical Properties ...... 364 C. Genomes ...... 365 xiv CONTENTS

D. Capsid Proteins ...... 368 E. Enzymes ...... 369 F. Antigenic Determinants ...... 370 III. Cell Specificity and Cultivation ...... 372 A. Differentiated Nature of Host Cells ...... 372 B. Cell-Culture Adaptation and the Role of Trypsin ...... 373 C. Adaptation by Reassortment ...... 375 D. Assay Systems ...... 375 E. Growth Kinetics ...... 377 F. Effects on Host Cells ...... 378 IV. Replication ...... 379 A. Adsorption ...... 379 B. Penetration and Uncoating ...... 379 C. Transcription ...... 380 D. Biosynthesis of Polypeptides ...... 380 E. Viral RNA Synthesis ...... 382 F. Assembly ...... 383 V. Genetics ...... 386 A. Isolation of Temperature-Sensitive Mutants ...... 386 B. Genome Reassortment ...... 387 C. Gene Coding Assignments ...... 387 VI. Biology of Rotaviruses ...... 389 A. Diseases and Pathogenesis ...... 389 B. Geographic and Host Range ...... 393 C. Epidemiology ...... 394 D. Strains and Serotypes ...... , ...... 399 E. Immunity ...... 401 F. Prospects for Control ...... 403 VII. Concluding Remarks ...... 405 References ...... 407

Chapter 9 Cytoplasmic Polyhedrosis Viruses Christopher C. Payne and Peter P. C. Mertens

I. Introduction ...... 425 II. Structural Properties of Polyhedra and Virus Particles ...... 427 A. Morphological and Biophysical Properties ...... 427 B. Structural Proteins ...... 435 C. RNA Components ...... 438 D. Cytoplasmic Polyhedrosis Virus Classification ...... 443 III. In Vitro Studies on the Mechanisms of Cytoplasmic Polyhedrosis Virus Replication ...... 452 A. Virus-Associated Enzymes ...... 453 CONTENTS xv

B. Translation of Cytoplasmic Polyhedrosis Virus RNA and Control of in Vitro Protein Synthesis ...... 470 IV. Cytoplasmic Polyhedrosis Virus Replication in Vivo ...... 476 A. Mode of Infection and Virus Morphogenesis ...... 477 B. Effect of Virus Infection on Cell and Host ...... 481 C. Cytoplasmic Polyhedrosis Virus Infection in Cell Cultures ...... 485 V. Cytoplasmic Polyhedrosis Virus Infections in Insect Populations ...... 487 A. Natural Incidence ...... 487 B. Cytoplasmic Polyhedrosis Viruses as Biological Control Agents ...... 489 VI. Conclusions ...... 491 References ...... 492

Chapter 10 The Plant Reoviridae R. I. B. Francki and Guido Boccardo

I. Introduction ...... 505 II. Members of the Reoviridae That Infect Plants ...... 506 A. Phytoreovirus ...... 506 B. Fijivirus ...... 507 C. Unclassified: Rice Ragged Stunt Virus ...... 510 D. Viruses Possibly Involved with Plant Diseases of Uncertain Etiology ...... 510 III. Properties of Virus Particles ...... 513 A. Virus Purification ...... 513 B. Virus Composition ...... 515 C. Virus-Particle Structure ...... 522 D. Antigenic Properties of the Viruses ...... 528 IV. Plant-Virus Relationships ...... 533 A. Diseases Caused by the Plant Reoviridae and Their Transmission ...... 533 B. Histopathology and Cytopathology of Infected Plants...... 540 V. Virus-Vector Relationships ...... 545 A. Virus Multiplication in Insects ...... 546 B. Virus Multiplication in Insect Cell Cultures ...... 547 VI. Classification of and Possible Relationships among the Reoviridae ...... 549 A. Classification Problems ...... 549 B. Possible Relationships among Existing Genera ...... 550 References ...... 553

Index ...... 565