Main description:

With the advent of genetic engineering methods and improved biochemical tech niques, much has been learned about the replication of influenza viruses, their structure and their epidemiology. It appears that the time is ripe to review these efforts and to provide a molecular perspective of influenza virology. It is hoped that this book will stimulate our thinking, help us in designing new experiments, and possibly show avenues leading to the control of the diseases associated with influenza viruses. Peter Palese, New York, N. Y. August 1983 David W. Kingsbury, Memphis, Tenn. Contents List of Contributors. . . . . . . . . . . . . . . . . XV 1. The Evolution of Influenza Viral Genetics - A Perspective. By E. D. Kilbourne. . . . . . . . . . . . . . . . 1 I. Introduction. . . . . . . . . . . . . . . . . 1 II. The Development of Modern Influenza Viral Genetics 2 A. Early Evidence of Genetic Variation in the Laboratory 2 B. Application of Formal Genetic Techniques to Studies of Influenza Virus . . . . . . . 3 C. Genetic Markers. . . . . . . . . 3 D. Development of Plaquing Systems. . . 4 E. The Use of Conditional Lethal Mutants 5 F. New Approaches in Influenza Virus Genetics. 6 1. The Biochemical Identification of Viral Gene Products in the Unambiguous Definition of Viral Inheritance . . . 6 2. Mapping of the Influenza Virus Genome by Correlative Physico-Chemical and Biological Techniques. . . . . . 7 3. The Application of Molecular Biological Techniques to the Study of Viral Genetic Variation. . . . . . . . . 8 4. Oligonucleotide Mapping of Viral RNA's . . . . . . . 8 5. Contribution of Protein and RNA Sequencing to Influenza Viral Genetics-Intragenic Mapping . . . . . . . 8 III. Viral Genetics and the Understanding of Viral Virulence and Pathogenicity . . . . . . . . . . . . . . . . . . .

Contents:

1. The Evolution of Influenza Viral Genetics - A Perspective.- I. Introduction.- II. The Development of Modern Influenza Viral Genetics.- A. Early Evidence of Genetic Variation in the Laboratory.- B. AppHcation of Formal Genetic Techniques to Studies of Influenza Virus.- C. Genetic Markers.- D. Development of Plaquing Systems.- E. The Use of Conditional Lethal Mutants.- F. New Approaches in Influenza Virus Genetics.- 1. The Biochemical Identification of Viral Gene Products in the Unambiguous Definition of Viral Inheritance.- 2. Mapping of the Influenza Virus Genome by Correlative Physico-Chemical and Biological Techniques.- 3. The Application of Molecular Biological Techniques to the Study of Viral Genetic Variation.- 4. Oligonucleotide Mapping of Viral RNA's.- 5. Contribution of Protein and RNA Sequencing to Influenza Viral Genetics-Intragenic Mapping.- III. Viral Genetics and the Understanding of Viral Virulence and Pathogenicity.- IV. Influenza Virus Genetics and the Epidemiology and Evolution of Influenza Viruses (Molecular Epidemiology).- A. Genetic Reassortment in Nature and Its Contribution to the Evolution of New Viruses.- B. The Genetics of Minor Variation.- C. Influenza A, B, and C Viruses.- V. Practical Applications of Influenza Viral Genetics.- VI. The Special Genetics of Divided Genome Viruses in Relation to Problems in Influenza.- VII. Unsolved Problems in Influenza and Genetic Approaches to Their Solution.- References.- 2. The Influenza Virus RNA Segments and Their Encoded Proteins.- I. Introduction.- II. The Influenza Virus Particle: Basic Structure.- III. Structure of the Genome.- A. Early Evidence for a Segmented Genome.- B. The 8 Segments of Influenza Virus RNA.- C. Methods for Assigning Gene Functions.- D. Sequences at the 5' and 3' Ends of Each RNA Segment Are Common.- E. Synthesis of Double-Stranded DNA from Influenza Vims RNA, Cloning and Nucleotide Sequencing.- IV. The Influenza Vims RNA Segments.- A. RNA Segments 1, 2, and 3: Properties of the Transcriptase Associated Proteins PBl, PB2, and PA.- B. RNA Segment 4: Stmcture and Function of the Hemagglutinin.- 1. The Stmcture of RNA Segment 4 Coding for the Hemagglutinin.- 2. The Three-Dimensional Stmcture of the Hemagglutinin.- 3. Synthesis of the Hemagglutinin, Cotranslational and Post- Translational Modifications.- 4. Cleavage Activation of Infectivity and in Vitro Fusion.- C. RNA Segment 5: The Stmcture of the Nucleocapsid Protein (NP).- D. RNA Segment 6: The Structure and Properties of the Neuraminidase.- E. RNA Segment 7: The Stmcture and Synthesis of the Membrane Protein (Mj) and Non-Structural Protein (M2).- F. RNA Segment 8: The Stmcture and Synthesis of Nonstmctural Proteins NSj and NS2.- G. Overlapping Coding Regions Using Different Reading Frames in Vimses.- Acknowledgements.- Appendix: The Influenza Vims Nucleotide Sequence (A/PR/8/34 Strain).- References.- 3. Transcription and Replication of Influenza Viruses.- I. Introduction.- II Viral mRNA Synthesis.- A. Priming by Cellular Capped RNAs-Discovery.- B. Priming by Cellular Capped RNAs-Mechanism.- C. Role of the Three Viral P Proteins in the Steps of Primed Transcription.- D. Termination of Transcription and Poly (A) Addition.- E. Regulation of Viral mRNA Synthesis in the Infected Cell.- F. Cellular Site of Viral mRNA Synthesis.- G. Role of Other Host Nuclear Functions in Viral mRNA Synthesis-Splicing and Methylation of Internal A Residues.- III. Synthesis of Full-Length Transcripts.- IV. Synthesis of vRNA (Replication).- References.- 4. Genetic Relatedness of Influenza Viruses (RNA and Protein).- I. Introduction.- II Genetic Relatedness of Viral RNAs.- A. Differences in Migration Rates of the RNA Segments As Revealed by Polyacrylamide Gel Electrophoresis (PAGE).- B. Molecular Hybridization.- 1. Direct RNA-RNA Hybridization.- 2. Competitive Hybridization.- 3. DNA-RNA Hybridization.- 4. Analysis of Double-Stranded Nuclease Si-Treated Hybrid Molecules by Polyacrylamide Gel Electrophoresis.- C. Oligonucleotide Fingerprints.- D. Sequencing of RNA Segments.- 1. Sequencing of 32P-End Labeled RNAs by Partial Nuclease Digestion.- 2. Sequencing of the 3' End of RNAs Using the Dideoxy Method.- 3. Sequencing of Total RNA Segments.- a) Hemagglutinin Gene.- b) Neuraminidase Gene.- c) The Three P-Protein Genes.- d) The Nucleoprotein Gene.- e) The Membrane Protein Gene.- f) The Nonstructural Protein Gene.- III. Genetic Relatedness of Viral Proteins.- A. Differences in Migration Rates of Viral Proteins As Revealed by Polyacrylamide Gel Electrophoresis.- B. Tryptic Peptide Mapping.- C. Direct Sequencing.- IV. Concluding Remarks.- Acknowledgements.- References.- 5. Antigenic Variation Among Type A Influenza Viruses.- I. Introduction.- II. Historical.- III. Nomenclature.- IV. Chemical and Physical Properties of the Antigens.- V. The Hemagglutinin (HA).- A. Isolation and Antigenic Properties of HA.- B. Changes in Conformation and Antigenicity of the HA at Low pH.- C. Antigenic Drift in the HA.- D. Use of Monoclonal Antibodies in the Analysis of Antigenic Drift.- E. Sequence Change in the HA of Influenza Virus Variants Selected with Monoclonal Antibodies.- F. Location of the Antigenic Sites in the 3-D Stmcture of the HA.- G. Sequential Selection of Antigenic Variants.- H. Antigenic Drift in the HA of Influenza A Virus from Lower Animals.- VI. The Neuraminidase (NA).- A. Antigenic Variation in the Neuraminidase.- B. Selection of NA Variants with Monoclonal Antibodies.- C. Antigenic Drift in the Neuraminidases of Influenza A Viruses from Lower Animals.- D. Mechanism of Antigenic Drift.- VII. Antigenic Shift.- A. Evidence from Sequence Data.- B. Possible Mechanisms of Shifts in Human Strains.- C. Antigenic Shift in Influenza Viruses from Lower Animals.- VIII. Variation in the Nucleoprotein.- IX. Variation in the Matrix Protein.- X. Variation in the Nonstmctural Proteins.- XI. Variation in the Polymerase Proteins.- XII What of the Future?.- Acknowledgements.- References.- 6. Expression of Cloned Influenza Vims Genes.- I. Introduction.- II. Expression Vectors.- III. Expression of Influenza Virus Genes in E. coli.- A. Expression of HA.- B. Expression of NSl Protein.- IV. Expression of Influenza Vims Genes in Eukaryotic Cells.- A. Expression of HA in Simian Cells Using Recombinant SV40 Viral Vectors.- B. Preparation of the HA Genes.- C. Construction of Recombinant Genomes.- D. Introduction of Recombinant Genomes into Simian Cells and Production of Virus Stocks.- E. Analysis of the HA Expressed from the SV40-HA Recombinant Vectors.- F. Quantitation of the Amounts of HA Expressed from the Recombinant Genomes.- G. Effect of Intervening Sequences in the Recombinant HA Transcript on the Level of Expression of HA Protein.- H. Expression of the Matrix Gene in Simian Cells Using an SV40-M Recombinant Virus.- I. Transient Expression of Cloned Genes in COS-1 Cells.- J. Continuous Expression of HA from Genes Integrated into the Chromosomes of Eukaryotic Cells.- V. Analysis of the Expression of Mutant HA Proteins.- A. The Signal-Minus HA Is a Non-Glycosylated, Intracellular Protein.- B. Removal of the C-Terminal Hydrophobic Sequence Converts HA into a Secreted Protein.- C. Future Prospects.- References.- 7. Mutants of Influenza Virus.- I. Introduction.- II. Characterization of Vims Mutants.- A. Influenza Vims-Cell Systems.- B. Nature of the Vims Population.- C. Natural Mutation Rate.- D. Induction of Mutants.- 1. Mutagens.- E. Leak and Reversion.- III. Temperature-Sensitive Mutants.- A. Genetic Interaction and Classification of Mutants.- B. Assignment of ts Lesions to Individual Genome Segments.- 1. Cambridge Mutants.- 2. New York-Tokyo Mutants.- 3. Giessen Mutants.- 4. Moscow Mutants.- 5. Bethesda Mutants.- 6. Summary.- C. Phenotypic Analysis of Temperature Sensitive Mutants.- 1. Assigment of Polypeptides to RNA Segments.- 2. Mutants with ts Lesions in RNA Segment 1.- a) Virion RNA Transcriptase Activity.- b) RNA-Dependent RNA Polymerase Activity of Infected Cell Extracts.- c) Virus-Specific RNA Synthesis in vivo.- d) Virus-Specific Polypeptide Synthesis.- e) Summary.- 3. Mutants with ts Lesions in RNA Segment 2.- a) Virion Transcriptase Activity.- b) RNA-Dependent RNA Polymerase Activity of Infected Cell Extracts.- c) Virus-Specific RNA Synthesis in vivo.- d) Virus-Specific Polypeptide Synthesis.- e) Summary.- 4. Mutants with ts Lesions in RNA Segment 3.- a) Virion Transcriptase Activity.- b) RNA-Dependent RNA Polymerase Activity of Infected Cell Extracts.- c) Virus-Specific RNA Synthesis in vivo.- d) Virus-Specific Polypeptide Synthesis.- e) Summary.- 5. Mutants with ts Lesions in RNA Segment 4.- 6. Mutants with ts Lesions in the RNA Segments Encoding the Nucleoprotein.- a) RNA Transcriptase Activity.- b) Virus-Specific RNA Synthesis in vivo.- c) Vims-Specific Polypeptide Synthesis.- d) Summary.- 7. Mutants with ts Lesions in the RNA Segment Encoding the Neuraminidase.- 8. Mutants with ts Lesions in RNA Segment 7.- 9. Mutants with ts Lesions in RNA Segment 8.- D. Temperature-Sensitive Mutant-Derived Vaccines.- IV. Cold-Adapted Mutants.- V. Host Range Mutants.- A. Non-Conditional Host Range Mutants.- B. Conditional Host Range Mutants.- C. Conclusions.- VI. Amantadine-Resistant Mutants.- VII. General Conclusions.- Acknowledgements.- References.- 8. The Stmcture of Influenza Virus Defective Interfering (DI) RNAs and Their Progenitor Genes.- I. Introduction.- II. Properties of Influenza DI Particles.- III. RNA of Influenza Vims DI Particles.- IV. Stmcture of Polymerase Genes and Polymerase Proteins.- A. Nucleotide Sequence of Polymerase Genes.- B. Primary Stmcture of PBl and PB2 Proteins.- C. Predicted Secondary Stmctures of PBl and PB2 Proteins.- V. Stmcture of DI RNAs.- A. Classes of DI RNAs.- B. Complete Nucleotide Sequences of DI RNAs.- VI. Generation of DI RNAs from the Progenitor RNAs.- VII. Transcription of Influenza Vims DI Vimses.- VIII. Interference by Influenza DI Viruses.- IX. Conclusion.- Acknowledgement.- References.- 9. Influenza B and Influenza C Viruses.- I. Introduction.- II. Influenza B Vims.- A. Influenza B RNA Species.- B. Influenza B Proteins.- 1. Hemagglutinin.- 2. Neuraminidase.- 3. Matrix Protein.- 4. The Non-Stmctural Proteins.- 5. The P Proteins.- C. Replication of Influenza B Vims.- D. Epidemiology.- E. Antigenic Variation in Influenza B Vimses.- III. Influenza C Vims.- A. Vims Stmcture.- 1. Morphology of Virions.- 2. Viral RNA Species.- 3. Viral Polypeptides.- 4. Fusion and Hemolysis Activities.- 5. Receptor Destroying Enzyme.- B. Influenza C Virus Replication.- C. Genetics and Epidemiology.- Acknowledgements.- References.- 10. Virus-Determined Differences in the Pathogenesis of Influenza Virus Infections.- I. Introduction.- II. Methods.- III. Hemagglutinin-Related Differences in Biologic Properties.- IV. Virulence in Chickens.- V. Neurovirulence in Mice.- VI. Virulence in Other Experimental Animals.- A. Ferrets.- B. Rats.- C. Mice.- VII Vims-Determined Differences in Other Systems.- A. Cell Culture.- B. Yield in Embryonated Eggs.- C. Sensitivity to Amantadine.- VIII. Virulence in Man.- IX. Summary.- References.- 11. Molecular Epidemiology of Influenza Virus.- I. Introduction.- II. Influenza A, B, and C Viruses and Associated Diseases.- III. Influenza in the 20th Century.- IV. Surveillance of Influenza Vimses.- V. The 1977 H1N1 Vimses.- VI. Molecular Epidemiology of Influenza in Animals.- VII. Mechanisms Contributing to Variation in Influenza Virus Field Strains.- A. Point Mutations.- B. Recombination (Reassortment).- C. Deletions/Insertions.- D. Recycling of Genes.- VIII. Outiook for the Future.- References.