Molecular Biology of the SARS-Coronavirus

1. Auflage 2010
ISBN: 978-3-642-03683-5
Verlag: Springer
Format: PDF
Kopierschutz: Adobe DRM (»Systemvoraussetzungen)

E-Book, Englisch, 328 Seiten

SARS was the ?rst new plague of the twenty-?rst century. Within months, it spread worldwide from its 'birthplace' in Guangdong Province, China, affecting over 8,000 people in 25 countries and territories across ?ve continents. SARS exposed the vulnerability of our modern globalised world to the spread of a new emerging infection. SARS (or a similar new emerging disease) could neither have spread so rapidly nor had such a great global impact even 50 years ago, and arguably, it was itself a product of our global inter-connectedness. Increasing af?uence and a demand for wild-game as exotic food led to the development of large trade of live animal and game animal markets where many species of wild and domestic animals were co-housed, providing the ideal opportunities for inter-species tra- mission of viruses and other microbes. Once such a virus jumped species and attacked humans, the increased human mobility allowed the virus the opportunity for rapid spread. An infected patient from Guangdong who stayed for one day at a hotel in Hong Kong led to the transmission of the disease to 16 other guests who travelled on to seed outbreaks of the disease in Toronto, Singapore, and Vietnam, as well as within Hong Kong itself. The virus exploited the practices used in modern intensive care of patients with severe respiratory disease and the weakness in infection control practices within our health care systems to cause outbreaks within hospitals, further amplifying the spread of the disease. Health-care itself has become a two-edged sword.

Autoren/Hrsg.

Inhaltsverzeichnis

1;Foreword;5
2;Preface;7
3;Contents;10
4;Part I: Viral Entry;13
4.1;Chapter 1: Cellular Entry of the SARS Coronavirus: Implications for Transmission, Pathogenicity and Antiviral Strategies;14
4.1.1;Introduction;14
4.1.2;The Spike Protein: Key to the Host Cell;15
4.1.3;The Attachment Factors DC-SIGN and DC-SIGNR: Enhancers or Inhibitors of SARS-CoV Infection?;17
4.1.4;The Two Faces of ACE2: SARS-CoV Receptor and Protector Against Lung Damage;18
4.1.4.1;The Structure of the Interface Between SARS-S and ACE2;19
4.1.4.2;Sequence Variations at the SARS-S/ACE2 Interface Might Impact Viral Transmission and Pathogenicity;20
4.1.4.3;The Human Coronavirus NL63 Uses ACE2 for Cellular Entry;21
4.1.4.4;SARS Versus NL63: A Correlation Between ACE2 Downregulation and Viral Pathogenicity?;21
4.1.5;Cleavage by Endosomal Cathepsin Proteases Activates SARS-S;23
4.1.6;Membrane Fusion is Driven by Conserved Elements Located in the S2 Subunit of the SARS Spike Protein;24
4.1.7;Conclusions;26
4.1.8;References;26
4.2;Chapter 2: The Cell Biology of the SARS Coronavirus Receptor, Angiotensin-Converting Enzyme 2;34
4.2.1;Introduction;34
4.2.2;Clues from Homologous Proteins;35
4.2.3;Regulation of ACE2 Expression on the Cell Surface;37
4.2.3.1;Proteolytic Cleavage Secretion;37
4.2.3.2;The Role of Membrane Microdomains;38
4.2.4;Conclusions and Future Perspectives;39
4.2.5;References;39
4.3;Chapter 3: Structural Molecular Insights into SARS Coronavirus Cellular Attachment, Entry and Morphogenesis;42
4.3.1;Structure of SARS Coronavirus (SARS-CoV);43
4.3.2;Structure of the Coronavirus Spike;45
4.3.3;Viral Membrane Fusion in SARS-CoV;47
4.3.4;Cellular Attachment and Entry of SARS-CoV;47
4.3.5;References;52
5;Part II: Structures Involved in Viral Replication and Gene Expression;55
5.1;Chapter 4: RNA Higher-Order Structures Within the Coronavirus 5 and 3 Untranslated Regions and Their Roles in Viral Replica;56
5.1.1;Introduction;56
5.1.2;cis-Acting RNA Elements in Coronavirus Replication;57
5.1.2.1;The Transcription Regulatory Sequence;58
5.1.2.2;The 5 cis-Acting RNA Elements;59
5.1.2.3;The 3 cis-Acting RNA Elements;62
5.1.2.4;Proteins Binding to the 5 and 3 cis-Acting Elements;64
5.1.3;Future Directions;66
5.1.4;References;66
5.2;Chapter 5: Programmed -1 Ribosomal Frameshifting in SARS Coronavirus;71
5.2.1;Introduction;71
5.2.2;Programmed -1 Ribosomal Frameshifting;72
5.2.3;Programmed Frameshifting Rates and Virus Propagation;73
5.2.4;Different Models, Different Assay Systems, Different Results;74
5.2.5;The Biology of -1 PRF in SARS-CoV is Different;75
5.2.6;A Unique Feature of the SARS-CoV Frameshift Signal: A Three-Stemmed mRNA Pseudoknot;76
5.2.7;A Second PRF Signal in SARS-CoV?;77
5.2.8;Summary and Perspectives;78
5.2.9;References;78
6;Part III: Viral Proteins;81
6.1;Chapter 6: Expression and Functions of SARS Coronavirus Replicative Proteins;82
6.1.1;Introduction;82
6.1.2;Organization and Expression of the Coronavirus Replicase Gene;85
6.1.3;Functions and Activities of Replicase Gene-Encoded Nonstructural Proteins;86
6.1.3.1;ORF1a-Encoded Nonstructural Proteins 1-11;86
6.1.3.2;ORF1b-Encoded Nonstructural Proteins 12-16;93
6.1.4;Future Directions;97
6.1.5;References;99
6.2;Chapter 7: SARS Coronavirus Replicative Enzymes: Structures and Mechanisms;106
6.2.1;The ADP-Ribose-1-Phosphatase Domain;107
6.2.2;The nsp7-nsp8-nsp9-nsp10 Cistron;109
6.2.2.1;The nsp7 Protein;109
6.2.2.2;The nsp8 Protein;110
6.2.2.3;The nsp9 Protein;113
6.2.2.4;The nsp10 Protein;115
6.2.3;The Endoribonuclease, nsp15;116
6.2.4;Conclusion;118
6.2.5;References;119
6.3;Chapter 8: Quaternary Structure of the SARS Coronavirus Main Protease;122
6.3.1;Introduction;123
6.3.2;Molecular Biology of the SARS-CoV Polyproteins;123
6.3.3;Structure of the SARS-CoV Main Protease;124
6.3.3.1;Three-Dimensional Structure of the SARS-CoV Main Protease;124
6.3.3.2;Quaternary StructureMproQuaternary structure of the SARS-CoV Main Protease;126
6.3.4;Enzyme Activity-AssayMproactivity-assay for the SARS-CoV Main Protease;126
6.3.5;Catalytic MechanismMproCatalytic mechanism of the SARS-CoV Main Protease;129
6.3.6;Structure and Function of the SARS-CoV Papain-Like Protease;131
6.3.7;Conclusions;131
6.3.8;References;133
6.4;Chapter 9: The Nucleocapsid Protein of the SARS Coronavirus: Structure, Function and Therapeutic Potential;136
6.4.1;Introduction;136
6.4.2;N-Protein: Structure and Composition;137
6.4.3;Stability of the N-Protein;138
6.4.4;Posttranslational Modification;138
6.4.5;Localization of the N-Protein;140
6.4.6;Genome Encapsidation: Primary Function of a Viral Capsid Protein;140
6.4.6.1;Recognition and Binding with the Genomic RNA;141
6.4.6.2;Formation of the Capsid;141
6.4.7;Perturbation of Host Cellular Process by the N-Protein;142
6.4.7.1;Deregulation of Host Cell Cycle;142
6.4.7.2;Inhibition of Host Cell Cytokinesis;143
6.4.7.3;Inhibition of Host Cell Translation Machinery;144
6.4.7.4;Inhibition of Interferon Production;144
6.4.7.5;Modulation of TGFbeta Signaling Pathway;144
6.4.7.6;Upregulation of COX2 Production;145
6.4.7.7;Upregulation of AP1 Activity;146
6.4.7.8;Induction of Apoptosis;146
6.4.7.9;Upregulation of Prothrombinase (hfgl2) Gene Transcription;147
6.4.7.10;Association with Host Cell Proteins;147
6.4.8;N-Protein: An Efficient Diagnostic Tool;148
6.4.9;N-Protein: A Suitable Vaccine Candidate;150
6.4.10;Future Perspective;151
6.4.11;References;153
6.5;Chapter 10: SARS Coronavirus Accessory Gene Expression and Function;159
6.5.1;Introduction;159
6.5.2;SARS-CoV Accessory Gene Expression and Function;162
6.5.2.1;ORF3a and ORF3b;162
6.5.2.2;ORF6;163
6.5.2.3;ORF7a and ORF7b;164
6.5.2.4;ORF8a and ORF8b;166
6.5.2.5;ORF9b;168
6.5.3;Conclusions;168
6.5.4;References;169
6.6;Chapter 11: SARS Accessory Proteins ORF3a and 9b and Their Functional Analysis;173
6.6.1;The SARS-CoV ORF3a Protein;173
6.6.2;Functional Analysis of ORF3a;175
6.6.3;Ion-Channel Activity of the ORF3a Homologue in Other Human Coronavirus;176
6.6.4;The SARS-CoV ORF9b Protein;178
6.6.5;Functional Role of ORF9b;179
6.6.6;SARS-CoV Accessory Proteins: An Important Part of the Virus Genome;179
6.6.7;References;180
6.7;Chapter 12: Molecular and Biochemical Characterization of the SARS-CoV Accessory Proteins ORF8a, ORF8b and ORF8ab;182
6.7.1;Introduction;182
6.7.2;Genetic Variations in the ORF8 Region;184
6.7.3;Expression of ORF8a, ORF8b and ORF8ab During Infection;185
6.7.3.1;Expression During Infection In Vivo;185
6.7.3.2;Expression During Infection In Vitro;186
6.7.4;Cellular Localization and Posttranslational Modification of ORF8a, ORF8b and ORF8ab;187
6.7.4.1;Cellular Localization;187
6.7.4.2;Glycosylation;187
6.7.4.3;Ubiquitination;188
6.7.5;Participation of ORF8a, ORF8b and ORF8ab in Viral-Viral Interactions and Their Effects on Other SARS-CoV Proteins;188
6.7.5.1;Interaction of ORF8a, ORF8b and ORF8ab with Other Viral Proteins;188
6.7.5.2;Downregulation of the E Protein by ORF8b;189
6.7.6;Impact of ORF8a, ORF8b and ORF8ab on Viral Replication and/or Pathogenesis;190
6.7.6.1;Contribution to Viral Replication;190
6.7.6.2;Abilities to Modulate Cellular Events;191
6.7.7;Conclusion;192
6.7.8;References;192
7;Part IV: Viral Pathogenesis and Host Immune Response;197
7.1;Chapter 13: SARS Coronavirus Pathogenesis and Therapeutic Treatment Design;198
7.1.1;Introduction;199
7.1.2;Human SARS-CoV Pathogenesis;199
7.1.2.1;The Clinical Course of Human SARS-CoV Infection;199
7.1.2.2;The Human Adaptive Immune Response to SARS-CoV;202
7.1.2.3;The Human Innate Immune Response to SARS-CoV;203
7.1.2.4;Animal Models of SARS-CoV Pathogenesis;203
7.1.2.5;In Vitro Models of SARS-CoV Pahtogenesis;207
7.1.2.6;SARS-CoV and MyD88;208
7.1.2.7;SARS-CoV and the Renin-Angiotensin System;210
7.1.3;SARS-CoV Therapeutic Design;213
7.1.3.1;SARS-CoV Antigenic Variation and Therapy Efficacy;213
7.1.3.2;Animal Models to Assess Passive Immunization Therapy Efficacy Against Divergent SARS-CoV Antigens;214
7.1.3.3;Animal Models to Assess Vaccine Immunization Therapy Efficacy Against Divergent SARS-CoV Antigens;215
7.1.3.4;SARS-CoV Vaccine Efficacy in Immunosenescent Populations;216
7.1.3.5;SARS-CoV Vaccine Immunopotentiation;220
7.1.4;Conclusion;222
7.1.5;References;223
7.2;Chapter 14: Modulation of Host Cell Death by SARS Coronavirus Proteins;234
7.2.1;Cell Death During SARS-CoV Infection;234
7.2.2;Induction of Host Cell Death by SARS-CoV Structural Proteins;236
7.2.2.1;E Protein;237
7.2.2.2;M Protein;237
7.2.2.3;S Protein;238
7.2.2.4;N Protein;238
7.2.3;Accessory Proteins;239
7.2.3.1;Protein 3a;241
7.2.3.2;Protein 3b;241
7.2.3.3;Protein 6;242
7.2.3.4;Protein 7a;242
7.2.3.5;Proteins 8a/8b;243
7.2.4;Conclusion;244
7.2.5;References;244
7.3;Chapter 15: SARS Coronavirus and Lung Fibrosis;249
7.3.1;Introduction;249
7.3.2;SARS-CoV-Mediated Lung Fibrosis;251
7.3.3;TGF-beta and SARS-Induced Lung Fibrosis;252
7.3.3.1;TGF-beta Signal Transduction;252
7.3.3.2;TGF-beta and Lung Fibrosis;252
7.3.3.3;The Role of TGF-beta in SARS-Induced Lung Fibrosis;253
7.3.4;ACE2/Angiotensin II and SARS-Induced Lung Fibrosis;254
7.3.5;Other Mechanisms of SARS-Mediated Lung Fibrosis;255
7.3.6;Conclusion;256
7.3.7;References;256
7.4;Chapter 16: Host Immune Responses to SARS Coronavirus in Humans;261
7.4.1;Introduction;262
7.4.2;Immune Responses;265
7.4.3;Innate Immunity;265
7.4.4;Viral Evasive Strategies;267
7.4.5;T Cell Responses;268
7.4.6;Antibody and B-Cell Responses;271
7.4.7;Vaccines;272
7.4.8;Summary and Perspectives;273
7.4.9;References;274
7.5;Chapter 17: The Use of Retroviral Pseudotypes for the Measurement of Antibody Responses to SARS Coronavirus;281
7.5.1;Introduction;281
7.5.2;Construction of Retroviral Pseudotypes Bearing SARS-CoV (S) Glycoproteins;282
7.5.3;Neutralization Assay;284
7.5.4;Expanding the Repertoire of SARS-CoV (S) Pseudotypes;284
7.5.5;Pseudotype Titration and Target Cell Lines;286
7.5.6;Reporter Systems;286
7.5.7;Correlation with Other Immunological Assays;287
7.5.8;Conclusion;288
7.5.9;References;288
7.6;Chapter 18: SARS Coronavirus Spike Protein Expression in HL-CZ Human Promonocytic Cells: Monoclonal Antibody and Cellular Trans;291
7.6.1;SARS Coronavirus: A Lethal Zoonotic Virus with Future Potential for Reemergence;292
7.6.2;SARS-CoV Spike Glycoprotein Contributes to Virulence and Pathogenesis;292
7.6.3;Generation of Human Monoclonal Antibodies by Selection from Recombinant Single-Chain Antibody-Phage Libraries Constructed from;293
7.6.4;Expression of SARS-CoV Spike Protein in Mammalian Cell Lines and Specific Detection by Human Monoclonal Antibodies: Potential ;294
7.6.4.1;Western Blot Characterization of HL-CZ Cells Transfected with Codon-Optimized SARS-CoV Spike Construct;294
7.6.4.2;Microscopic Analysis of SARS-CoV Spike Expression in HL-CZ Cells by Indirect Immunofluorescence Labeling;295
7.6.5;Transcriptomic Analysis Reveals Differentially Regulated Genes in HL-CZ Cells Following SARS-CoV Spike Transfection;297
7.6.5.1;Immune and Inflammatory Genes;298
7.6.5.2;Cell Cycle Genes;301
7.6.5.3;Cytoskeletal Genes;302
7.6.5.4;Trafficking and Transport Genes;303
7.6.6;Conclusions and Future Prospects;303
7.6.7;References;304
7.7;Chapter 19: Signaling Pathways of SARS-CoV In Vitro and In Vivo;307
7.7.1;Introduction;307
7.7.2;p38 MAPK Signaling Pathway in Viral Infection;309
7.7.3;p38 MAPK Signaling Pathway in SARS-CoV-Infected Cells;311
7.7.4;Downstream Signaling Pathways of p38 MAPK;312
7.7.5;ERK1/2 Activation by SARS-CoV Infection;313
7.7.6;JNK Activation by SARS-CoV;313
7.7.7;PI3K/Akt Activation by SARS-CoV;314
7.7.8;NF-kappaB Activation and Inhibition by SARS-CoV Proteins;315
7.7.9;Inhibitory Effects of Viral Proteins on the Cell Cycle;315
7.7.10;Apoptotic Signaling Pathway;316
7.7.11;Signaling Pathways in SARS Patients;317
7.7.12;Conclusion;318
7.7.13;References;319
8;Index;325