Reoviruses. Virion. Genome. Genes and proteins. Viruses and hosts. Diseases. Distinctive characteristics
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1 Reoviruses Virion Genome Genes and proteins Viruses and hosts Diseases Distinctive characteristics
2 Virion Naked icosahedral capsid (T=13), diameter nm Capsid consists of two or three concentric protein shells Inner capsid (T=1), or core, contains RNA pol and capping enzymes
3 Genome Linear double-stranded RNA, segments Total genome length kb
4 Genes and proteins mrnas are full length copies of each genome segment Typically one protein is encoded per genome segment 6-8 capsid proteins 3-6 nonstructural proteins
5 Viruses and hosts Reoviridae from respiratory enteric orphan viruses 12 genera, including Orthoreovirus, Rotavirus, Orbivirus Infects humans (reoviruses types 1-3, rotaviruses, and Colorado tick fever virus), mammals, birds, fish, mollusks, plants, insects, and fungi
6 Viruses and hosts
7 Diseases Members of genus Orthoreovirus cause little or no disease in humans Rotavirus is an important cause of gastroenteritis worldwide and infant mortality in the developing world Viruses spread between hosts by direct transmission, contaminated food or water, or arthropod vectors
8 Reoviruses were the first double-stranded RNA viruses discovered Reoviruses caused inclusion bodies in infected cells that contained double stranded RNA Some members of the Reoviridae are important pathogens
9 Genome Reoviridae have segmented genomes made of double-stranded RNA discrete linear segments of dsrna Three large (L) genes, three medium (M) genes and four small (S) genes Mixed infections with different virus strains can result in reassortant viruses
10 Generation of reassortant viruses
11 Genome General characteristics of a mammalian reovirus gene segment
12 Virion Reovirus virions contain concentric layers of capsid proteins Inner capsid (T=1) assembled from major (λ1 and σ2), and minor (µ2 and λ3) proteins Outer capsid (T=13) constructed from complexes of µ1 and σ3 and a spike made from pentamers of λ2
13 The attachment protein binds to one or two cellular receptors Sialic acid Junctional adhesion molecule A (JAM-A) A transmemebrane protein located at tight junctions
14 During entry, the outer capsid is stripped from virions and the core is released into the cytoplasm Entry is by clathrin-mediated endocytosis dependent on β1 integrin
15 In endosomes, virions undergo partial degradation by cathepsins to form infectious subvirion particles (ISVPs) Disassembly can be viewed using cryoelectron microscopy
16 Enzymes in the viral core synthesize and cap messenger RNAs All 10 mrnas are synthesized at the same time and passed up into the spike turret
17 Translation of reovirus mrnas is regulated Proteins made from long mrnas are made in lower abundance than those made from short mrnas
18 Interferon and PKR: effects on viral and cellular protein synthesis Reovirus dsrna is a potent inducer of: interferons which activate double-stranded RNA-dependent protein kinase R 2, 5 -oligo(a) synthetase which activates ribonuclease L
19 Synthesis of progeny double-stranded genomes occurs within subviral particles RNA replication and virion assembly take place in intracytoplasmic inclusions called virus factories
20 mrna bound to non-structural proteins and outer capsid proteins represent the first step in assembly Replicase particles then form which are engaged in (-) strand synthesis Secondary transcriptase particles then make (+) strand RNA Outer capsids are formed when sufficient capsid proteins have been synthesized
21 Assortment of gene segments and virion assembly
22 Reoviruses induce apoptosis via activation of innate immune response transcription factors NF-κB and IRF-3 Caspases are also activated Cell cycle is arrested by inactivation of cyclindependent kinases (CDK)
23 Studies of reovirus pathogenesis in mice Best characterized model is reovirus pathogenesis in mouse central nervous system Infection of ependymal cells lead to nonlethal hydrocephalus Type 3 reoviruses infect neurons causing encephalitis
24 Reovirus also infects liver, bile duct and can cause myocarditis in mice Infection of bile ducts causes ductal obliteration and jaundice Attachment protein σ1 is responsible for cell tropism
25 Plasmid-based reverse genetics for Reovirus
26 Fundamental concepts Reoviridae have double or triple layered capsids with icosahedral symmetry, and form different doublestranded RNA genome segments Rotaviruses cause severe gastroenteritis and are a major cause of disease and death in children in the developing world During entry into endosomes, reoviruses lose their outer capsid shell, and the cores penetrate the endosomal membrane and enter the cytoplasm Reovirus cores contain viral RNA polymerases, which make viral mrnas by transcribing each of the up to 12 viral genome segments
27 Fundamental concepts Viral mrnas are capped by viral capping enzymes during transcription, and are extruded from the core into the cytoplasm The reovirus outer capsid protein binds to dsrna, reducing activation of PKR and therefore allowing viral protein synthesis to continue Virus factories are localized areas of the cytoplasm where viral RNA synthesis and virion assembly take place Viral mrnas are selectively incorporated into replicase particles, where the viral RNA pol copies them
28 Fundamental concepts The resulting virus cores can either continue to synthesize viral mrnas or can be further encapsidated to make mature reovirus virions Reoviruses have been used to study aspects of viral pathogenesis in mice
29 Key terms 2,5 -oligo(a) synthetase Apoptosis Caspases Cathepsins Cryoelectron microscopy Cyclin-dependent kinase (CDK) Double-stranded RNAdependent protein kinase (PKR) Encephalitis Ependymal cells Hydrocephalus Inclusion bodies Infectious subvirion particles (ISVPs) Interferons Jaundice Junctional adhesion molecules (JAM-A) Myocarditis NF-κB (nuclear factor κb) Ras Reassortant viruses Ribonuclease L Sialic acid Tight junction tropism
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