The Molecular Biology of Poliovirus

602 Pages·1985·39.285 MB·English

by  Dr. Friedrich Koch, Prof. Dr. Gebhard Koch (auth.)

The Molecular Biology
ofP oliovirus
Friedrich Koch and Gebhard Koch
Wien New York
Springer-~rlag
Dr. Friedrich Koch
Prof. Dr. Gebhard Koch
Abteilung Molekularbiologie
Universitat Hamburg, Federal Republic of Germany
This work is subject to copyright.
All rights are reserved, whether the whole or part of the material is concerned, specifically those
of translation, reprinting, re-use of illustrations, broadcasting, reproduction by photocopying
machine or similar means, and storage in data banks.
© 1985 by Springer-VerlaglWien
Softcover reprint of the hardcover 1st edition 1985
With 122 Figures
Library of Congress Cataloging in Publication Data. Koch, Friedrich, 1957-. The molecular biology of poliovirus.
Includes index.!. Poliovirus. 2. Molecular biology. I. Koch, Gebhard. II. Title. QR201. P73K58. 1985. 576'.6484. 85-4757.
ISBN-13:978-3-7091-7467-8 e-ISBN-13:978-3-7091-7000-7
DOl: 10.1007/978-3-7091-7000-7
Preface
Years ago when we were asked to write a book on the present-day knowledge of
the molecular biology of poliovirus, we did not expect that such an apparently
simple task could involve so much time and effort. Our writing was hampered by
the fact that both of us are full time "workers", so that this monograph is mainly a
spare time expedience.
The main attention of this book focuses on a detailed review of the molecular
biology of poliovirus and especially on the advances of the last decade; medical
and environmental aspects are only briefly mentioned. Observations from older
studies are considered in view of more recent information. Some of the older ob
servations provided fundamental insights and paved the way for present day
research; too often such data has been neglected or independently rediscovered.
Today, poliovirus research has again attracted considerable interest. High points
gained within the last few years were the elucidation of the complete nucleotide
sequences of the RNAs of the three poliovinls serotypes and the corresponding
vaccine strains, the demonstration of genome evolution during transmission of
poliovirus in an epidemic, further characterization of the antigenic sites on the
virus particle and of the antigenic drift, characterization of alternate conforma
tional states of the virion capsid, the development of monoclonal antibodies
against some of the virus proteins, observations on the role of the plasma
membrane, cytoskeleton, and cytoplasmic membranes as mediators in the virus
induced redirection of the synthetic machinery of the host cell, and characteriza
tion of proteins involved in RNA replication. The review is certainly not all
inclusive. We are grateful for any comments, corrections, additions, and criticism.
We wish to thank Kurt Bienz, John Bilello, Richard Crowell, Keith Dunker,
Denise Egger, John Mapoles, Paolo La Colla, Roland Rueckert, and Douglas
Scraba for many fruitful discussions and helpful comments. We also thank all
those who provided us with material for illustrations.
The personal proceeds of this book will be donated to a preventive health care
project in Nicaragua. As part of this project, the Nicaraguan health ministry has
initiated mass vaccination programs against poliomyelitis, which will for the first
time succeed in reaching most of the children in this country.
Hamburg, March 1985 F. Koch and G. Koch
Contents
Part I: The Poliovirus.
1. History. . . . . 3
I. Introduction . 3
II. Early History-Occasional Nonepidemic Cases of Poliomyelitis 4
III. The 18th and 19th Centuries: Polioepidemics, Poliomyelitis Is
Described as a Clinical Entity . . . . . . . . . . .. 6
IV. Early 20th Century: Research on Polio Begins, Virology Is still
a Clinical Discipline. . . . . . . . . . . . . . .. 6
V. The Development of Vaccines Against the Poliomyelitis Virus 9
VI. The Fifties and the Advent of Molecular Biology . . . 10
VII. Advances in Polio Research During the Past Two Decades 11
2. Classification. . . . . . . . 15
I. General Considerations. . 15
A. Nature of the Genome. 15
B. The Cryptogram . . . 19
II. The Distinguishing Features of Picornaviruses . 19
III. Comparison of Different Picornavirus Genera. 24
A. Disease Aspects 25
B. Serotypes. . . . . . . . . . 25
C. Physical Properties . . . . . . 25
D. Relative Relatedness to Polioviruses 26
3. Composition and Structure of the Virion 28
I. Introduction......... 28
II. Composition and Physical Properties of the Virion. 30
III. Overall Architecture of the Viral Capsid 38
A. Building Blocks . . . . . . . . 38
1. Functional Requirements. . . . 38
2. Biological Arguments of Building Block Economy and
Efficiency of Assembly. . . . . . . . . . .. 38
3. Thermodynamic Forces and Structural Consequences 38
B. The Geometric Design: The Icosahedron 39
1. Helical Tubes Versus Spherical Shells . . . . .. 39
VIII Contents
2. Cubic Symmetries, the Platonic Polyhedra 39
3. The Icosahedron. . . . . . . . . . 40
a) Advantages of Icosahedral Symmetry . 40
b) Limitations of the Icosahedral Skeleton as a Model for
Virus Structure . . . . . . . . . . . . . . 41
c) Related Regular Polyhedra with Icosahedral Symmetry 42
d) The Bonding Pattern of an Icosahedral Lattice. . . 42
C. Experimental Evidence that the Poliovirus Capsid Is an Ico-
sahedral Lattice . . . . . . . . 44
1. X-Ray Diffraction Studies. . . . . . . . . 45
2. Electron Microscopic Observations . . . . . 45
3. Resolving the 32 or 60 Capsomers Controversy 47
4. Similar Construction Principles for Picornaviruses and the
Small Plant Viruses. . . . . . . . . . . . . . 47
IV. Characterization of the Building Blocks: The Capsid Proteins. 50
A. Separation and Identification . . . . 50
B. Amino Acid Composition and Sequence . . . . . . . 53
C. Microheterogeneity . . . . . . . . . . . . . . . 59
V. Relative Localization of the Viral Proteins in the Capsid and
Bonds Involved. . . . . . 59
A. General Reflections. . . . . . . . . . . . . . . 59
1. Geometric Restrictions. . . . . . . . . . . . . 60
2. Structural Principles Borrowed from Plant Picornaviruses 64
3. Implications for the Capsid Features of Poliovirus: The
Concept of a Rigid Capsid Backbone and Variable Surface
Protrusions . . . . . . . . . . . . . .. 69
B. Experimental Results . . . . . . . . . . .. 71
1. Specific Chemical Modifications of Capsid Proteins 71
2. Chemical Crosslinking of Capsid Proteins. 75
3. UV Irradiation of Poliovirus. . . . . . . 78
VI. The Dissociation of Poliovirus . . . . . . . . 79
A. Breakdown During Preparatory Procedures for Electron
Microscopy. . . . 82
B. Alkaline Degradation 84
C. Heat Degradation. . 86
D. Guanidine Degradation. 89
E. Urea Degradation. . . 89
F. Reassociation of Poliovirus from Products of Urea Degrada-
tion. . . . . . . . . . . . . . . . 93
G. Conclusions. . . . . . . . . . . . . . . . . . 93
VII. Conformational Forms of the Poliovirus Capsid. . . . . . 94
A. Poliovirus Capsid Structures During Early Interactions with
the Host Cell and upon Experimentally Induced Disruption:
Two Conformational States of the Shell 95
B. Viral Structures During Maturation . . . . . . . . . 97
Contents IX
C. Two Distinct, Reversibly Interconvertible Conformational
States of Intact Poliovirions. . . . . . . . . . . . 99
D. Conformational Alterations Monitored by Changes in the
Intrinsic and Induced Fluorescence of Poliovirus Com
ponents . . . . . . . . . . . . . . . . . . . 103
E. Dense Particles. . . . . . . . . . . . . . . . . 106
F. Possible Functions of the Alternate Conformational States
of the Poliovirus Capsid . . . . . . . . . . . . . 108
VIII. Antigenicity. . . . . . . . . . . . . . . . . . . 114
A. The Main Poliovirus Antigenic Sites: The D-and C-Antigenic
States . . . . . . . . . . . . . . . . . . . . 118
1. Attempts to IdentifY the C-and D-Antigenic Determinants
in Terms of the Constituent Capsid Proteins with Poly-
clonal Antisera. . . . . . . . . . 121
2. Studies with Monoclonal Antibodies. . 126
B. Other, Minor Antigenic Sites on Poliovirus 128
C. Serodifferentiation of Polioviruses . 132
IX. Summary. . . 132
A. Composition 132
B. Structure. . 133
C. The Capsid Proteins. 133
1. Number of Peptides and Type of Association on the
Structure Units 133
2. Relative Localizations . . . . 134
3. Heterogeneity. . . . . . . 135
D. Two Conformational States of the Capsid. 135
E. Antigenicity. . . . . . . 136
4. Structure and Function of the Genome 138
I. Introduction..... 138
II. Genome Structure. . . . 139
A. Characteristic Features . 139
B. Structural Organization 140
C. Secondary Structure. . 142
D. The Genome-Linked Protein VPg . 143
III. Nucleotide Sequences . . . . 146
A. Oligonucleotide Mapping. . . . 146
B. Cloning of Poliovirus cDNA 150
C. The Consensus Sequence of Poliovirus Type I 150
D. Terminal Sequences. 161
1. 5' End Sequences 161
2. 3' End Sequences 165
3. The Poly (A) Tract 166
E. Features of the Coding Region. 167
1. Codon Usage . . . . . . 167
2. Cleavage Signals for Proteolytic Processing 167
x
Contents
IV. Genome Products and Gene Order. 171
A. Mapping of the Genome Products on the Polioviral
Genome .......... . 175
1. Relatedness of Viral Proteins: Tryptic Peptide Analysis 176
2. Biochemical Mapping . . . . . . . . . . . . 177
3. The Genome Map as Deduced from Genetic Studies 178
B. Function of the Viral Proteins. . . . . . . . . . 178
1. The P-1 Proteins: Coat-Proteins, Proteinkinase, Shut-Off of
Protein Synthesis. . . . . . . . . . . . . . . 180
2. The P-2 Proteins: Guanidine Sensitivity, VPO Protease . 180
3. The P-3 Proteins: glu/gly Protease, VPg Replicase, Cyto
phathic Effect. . . . . . 181
V. Genomic Variation of Polioviruses . 182
VI. Genetics . . . 188
A. Mutations . . . . . . . . 188
B. Mutant Types . . . . . . . 191
1. Temperature Sensitive Mutants. 191
2. Structural Markers . . 192
3. Non Structural Markers . 195
C. Genetic Recombination 195
D. Mechanism of Recombination 199
VII. Summary. . . . . . . . 200
Part II: The Replication of Poliovirus 203
5. Introduction . . . . . . . . . 203
I. The "Life-Cycle" of Poliovirus 203
II. Timecourse of Poliovirus Replication 208
III. The Host Cell . . . . . 212
A. Constituents of the Cell 213
B. The Nucleus . . . . 215
C. The Plasma Membrane. 215
D. The Ionic Environment 216
E. The Cytoskeleton. . . 218
F. Intracellular Membranes 220
G. The Cell Cycle 220
H. Employment of the Metabolic Machinery of the Host Cell
by the Virus . . . . . . . . . . . . . . 221
IV. Some Speculations on Abortive Infections of Poliovirus. . . 222
6. Morphological Alterations of the Host Cell as an Essential Basis for
Poliovirus Replication. . . . . 226
I. Introduction...... 226
II. Microscopic Oberservations 227
A. Light Microscopy. . 228
B. Electron Microscopy. . 230
Contents XI
1. The Nucleus 230
2. Ribosomes . 235
3. "Viroplasm". 235
4. Alterations of Intracellular Membranes . 237
a) Nuclear "Extrusions" . . . . . . 237
b) Membraneous Cisternae . . . . . 240
c) Biochemical Aspects of Membrane Formation 245
5. Changes in the Cytoskeletal Framework . 250
6. Assembly and Release of Progeny Virions. 250
7. Lysosomes and Autophagic Vesicles. . . 252
8. Inhibitors of Morphological Alterations 253
III. Speculations on the Function of Compartmentalization in Virion
RNA Synthesis and Assembly . . . . . . . . . . . . 256
A. Concerning the Mode of Vesicle Formation. . . . . . 256
B. Concerning the Relative Localization of RNA Synthesis and
Virion Assembly with Respect to the Vesicle Membrane 258
IV. Poliovirus induced Alterations in Functions of the Plasma
Membrane, in the Intracellular Ionic Environment, and in Cell
Size. . . . . . . . . . . . . . . . . . . . . . 262
A. Membrane Changes Accompanying Adsorption and Penetra-
tion of Poliovirus. . . . . . . . . . . . . . . . 262
B. Membrane Alterations Accompanying Virus Replication at
the Maximal Rate 264
V. Summary.......... 265
7. Early Interactions of Virus and Host Cell 267
I. Introduction......... 267
A. Overview and Definition of Terms 267
B. The Superposition of Abortive and Productive Pathways in
Infection. . . . . . 270
II. Adsorption and Attachment 271
A. The Reaction Partners . 272
1. The Virus Particle 272
2. The Virus Receptor Complex on the Host Cell 275
a) Properties of the Poliovirus Receptor . . 276
b) Other Functions and Components of the Receptor
Complex. . . . . . . . . . . . . 278
c) Number of Attachment Sites/Cell . . . . 278
d) Specificity and Genetics of Virus Receptors. . . . 279
B. The Interaction of Poliovirions with the Host Cell Mem-
brane . . . . 280
1. Adsorption . 282
2. Attachment . 285
XII Contents
3. Response of the Plasma Membrane . . . . . 286
a) Changes in Membrane Fluidity and Capping of
Viruses . . . . . . . . .. ..... 286
b) Changes in Membrane Permeability and Membrane
Potential. . . . . . .. .... 288
c) Interaction with Modifying and Stabilizing Mem
brane Components. . . . . . . . . . . . . 288
III. Penetration of Virus Particles into the Cell: Insertion and Phago-
cytosis . . . . . . . . . . . . . 291
IV. Uncoating: A Multistep Process. . . . . 297
A. Possible Steps and Sites of Uncoating . 297
B. The Fate of the Parental Capsid Proteins 300
V. Infection of Cells Lacking Receptors . 301
A. Introduction . . . . . . . . 301
B. Adsorption of Viral RNA to Cells 302
C. Penetration of Isolated Viral RNA into Cells. 304
1. RNA-Penetration by Passive Influx of RNA 304
2. Stimulation of Active Uptake of Viral RNA . 304
3. Entrance of Poliovirus RNA into Cells Via Lipid Vesicles
(Liposomes). . . . . . . . . . . . . . 305
D. Cellular Competence for Infection by Viral RNA. . . . 305
1. Optimal Conditions for the Use of Polycations. . . . 306
2. Relationship Between RNA Concentration and Yield of
Infections Centers . . . . . . . . . . . . . . 307
3. The Combined Effect of Dimethylsulfoxide and DEAE
Dextran on the Competence of Cells for Infection by
Viral RNA . . . . . . . . . . . . . . . . . 308
4. Competence of HeLa Cells for Infection by Viral RNA
at Different Stages in a Cell Growth Cycle 309
E. Conclusions. 309
VI. Summary. . . . . . . 310
8. Translation of the Viral Genome. 313
I. Mammalian Protein Synthesis . . . . 313
A. The Protein Synthesizing Machinery . 313
1. Ribosomes . . . . . . . . 314
a) Structure and Composition. . 314
b) Monosomes and Polysomes. . 314
c) Free and Membrane Bound Polysomes . 315
2. mRNA ........... . 315
a) The Cap. . . . . . . . . . . . 315
b) The 5' Terminal Untranslated Region . 317
c) The 3' Terminal Untranslated Region and the Poly
A Tract . . . . . . . . . . . . . . . 319