The ultrastructure of the developing replication site in foot-and-mouth disease virus-infected BHK-38 cells

Monaghan, Paul; Cook, H.; Jackson, Terry; Ryan, Martin D.; Wileman, Tom

doi:10.1099/vir.0.19408-0

Cited by 78 publications

(78 citation statements)

References 26 publications

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“…Most of the rhinovirus-induced vesicles appear to have single membranes, in contrast to poliovirus-induced vesicles. However, the formation of predominantly single-membraned vesicles has been reported for other picornaviruses (49), and vesicles of this morphology have also been observed during poliovirus infection (61).…”

Section: Discussionmentioning

confidence: 98%

Amino Acid Changes in Proteins 2B and 3A Mediate Rhinovirus Type 39 Growth in Mouse Cells

Harris

Racaniello

2005

J Virol

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Many steps of viral replication are dependent on the interaction of viral proteins with host cell components. To identify rhinovirus proteins involved in such interactions, human rhinovirus 39 (HRV39), a virus unable to replicate in mouse cells, was adapted to efficient growth in mouse cells producing the viral receptor ICAM-1 (ICAM-L cells). Amino acid changes were identified in the 2B and 3A proteins of the adapted virus, RV39/L. Changes in 2B were sufficient to permit viral growth in mouse cells; however, changes in both 2B and 3A were required for maximal viral RNA synthesis in mouse cells. Examination of infected HeLa cells by electron microscopy demonstrated that human rhinoviruses induced the formation of cytoplasmic membranous vesicles, similar to those observed in cells infected with other picornaviruses. Vesicles were also observed in the cytoplasm of HRV39-infected mouse cells despite the absence of viral RNA replication. Synthesis of picornaviral nonstructural proteins 2C, 2BC, and 3A is known to be required for formation of membranous vesicles. We suggest that productive HRV39 infection is blocked in ICAM-L cells at a step posttranslation and prior to the formation of a functional replication complex. The observation that changes in HRV39 2B and 3A proteins lead to viral growth in mouse cells suggests that one or both of these proteins interact with host cell proteins to promote viral replication.Picornaviruses are small RNA viruses with a single-stranded, positive-sense genome of approximately 7 to 8 kb enclosed in a nonenveloped, icosahedral capsid. These viruses are responsible for a wide range of diseases including foot-and-mouth disease, hepatitis, poliomyelitis, and the common cold. Following cell entry, the viral genome is translated as a single polyprotein that is co-and posttranslationally cleaved by viral proteases, yielding both precursor proteins and end products. Many of the precursor proteins are essential for virus replication, with functions distinct from the end products (29,40,42). Four structural proteins produced from the P1 region of the genome form the viral capsid. Nonstructural proteins of the P2 and P3 region of the genome encode all of the viral proteins responsible for genomic RNA replication.Viral genome replication occurs on the surface of cytoplasmic vesicles induced by the synthesis of viral proteins 2C and 3A and the precursor 2BC (70). These vesicles are formed in conjunction with massive cell membrane proliferation (4, 7) and increased cellular synthesis of phospholipids (32). With the exception of the two viral proteases 2A pro and 3C pro , all of the nonstructural proteins have been found on the surface of these vesicles and are thought to interact in the replication complex (26,59,61,70). The vesicles assemble into a rosette, and the replication complex forms on the inner face of the rosette (13, 26). Salt-induced dissociation of the rosette demonstrated that the individual vesicles themselves can also support replication initiation and elongation (26). Neither t...

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Section: Discussionmentioning

confidence: 98%

Amino Acid Changes in Proteins 2B and 3A Mediate Rhinovirus Type 39 Growth in Mouse Cells

Harris

Racaniello

2005

J Virol

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“…Induction of vesicular clustering and concentration of viral material for facilitated viral production is a well-known mechanism that operates in many positive-sense RNA viruses and poxviruses (Limpens et al, 2011;Monaghan et al, 2004;Welsch et al, 2009;Laliberte and Moss, 2010). Nevertheless, this hypothesis, as appealing as it might be, awaits compelling experimental support because, at the moment, the relevance of the clustered Rab11 vesicles is still uncertain.…”

Section: Discussionmentioning

confidence: 99%

Influenza A virus ribonucleoproteins modulate host recycling by competing with Rab11 effectors

Vale-Costa

Alenquer

Sousa

et al. 2016

Journal of Cell Science

127

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Influenza A virus assembly is an unclear process, whereby individual virion components form an infectious particle. The segmented nature of the influenza A genome imposes a problem to assembly because it requires packaging of eight distinct RNA particles (vRNPs). It also allows genome mixing from distinct parental strains, events associated with influenza pandemic outbreaks. It is important to public health to understand how segmented genomes assemble, a process that is dependent on the transport of components to assembly sites. Previously, it has been shown that vRNPs are carried by recycling endosome vesicles, resulting in a change of Rab11 distribution. Here, we describe that vRNP binding to recycling endosomes impairs recycling endosome function, by competing for Rab11 binding with family-interacting proteins, and that there is a causal relationship between Rab11 ability to recruit family-interacting proteins and Rab11 redistribution. This competition reduces recycling sorting at an unclear step, resulting in clustering of single- and double-membraned vesicles. These morphological changes in Rab11 membranes are indicative of alterations in protein and lipid homeostasis during infection. Vesicular clustering creates hotspots of the vRNPs that need to interact to form an infectious particle.Fundação para a Ciência e Tecnologia grants: (PTDC/IMI-MIC/1142/2012, SFRH/BPD/94204/2013, SFRH/BPD/62982/2009, IF/00899/2013); Fundação Calouste Gulbenkian; Instituto Gulbenkian de Ciência

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“…Lesion formation takes place rapidly within infected animals, but this is not too surprising as it has been reported that the cytopathic effect of FMDV can be detected in BHK-38 cells, which are a particularly susceptible tissue culture cell line, as soon as 1.5 h p.i. (17). While this is an extreme case, the doubling rate of FMDV in pigs has been estimated to around 2.3 h (Quan and Alexandersen, unpublished).…”

Section: Discussionmentioning

confidence: 99%

“…For sectioning, the method was essentially as described previously (17,18). The tissues were trimmed to an oblong shape if needed and then attached to the holder of a vibrating microtome (Leica Microsystems, Milton Keynes, United Kingdom) with superglue.…”

Section: Virusmentioning

confidence: 99%

Use of Confocal Immunofluorescence Microscopy To Localize Viral Nonstructural Proteins and Potential Sites of Replication in Pigs Experimentally Infected with Foot-and-Mouth Disease Virus

Monaghan

Simpson

Murphy

et al. 2005

J Virol

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Replication of foot-and-mouth disease virus in infected pig epithelium has been studied by immunofluorescence labeling of the viral nonstructural protein 3ABC and confocal microscopy. The results were correlated with viral RNA copy numbers in tissue samples from adjacent sites determined by reverse transcription-PCR (RT-PCR). Lesion formation was seen in the tongues and coronary band epithelia of infected pigs 2 days after infection. Viral replication was observed in cells of the epithelium of the tongue and coronary band but not in the associated stromal cells. Infected epithelial cells were present in the stratum spinosum, away from the lesion, with small lesions formed above the basement membrane. Viral replication was markedly reduced in tongue epithelium by day 3 postinfection but remained apparent in the coronary band tissue up to 5 days postinfection. These results were confirmed by the RNA copy number determined by RT-PCR.The causative agent of foot-and-mouth disease (FMD) is a highly infectious virus which can infect a wide range of clovenhoofed animals, including domestic cows, pigs, and sheep (3, 6). FMD virus (FMDV) is a member of the Picornaviridae and is a small (25-nm) nonenveloped virus with a genome consisting of a single strand of positive-sense RNA. Once the virus is inside a cell, the RNA is released from the capsid and translated into a single polyprotein which is cleaved into individual proteins. These consist of structural proteins that will form the viral capsid and nonstructural proteins which are involved in viral replication (8).Clinical signs of the disease vary in severity between species, with cows and pigs showing greater severity of signs than sheep (6). The most obvious effects of FMDV infection are elevated temperature and the appearance of lesions on the tongue and epidermis adjacent to the hoof, namely, the coronary band and, in ruminants, interdigital skin. Lesions first appear in animals which have been infected by contact with infected animals within 1 to 5 days of contact, depending on the intensity of contact, the virulence of the virus, and the dose used to inoculate the pigs. Inoculated animals have viremia from around 1 day after inoculation, and it is assumed that the spread of the virus within such infected animals is by the general circulation.The precise nature of the virus-cell interactions that give cell type specificity in vivo is not certain, although host cell integrins have been proposed as receptors for FMDV (11)(12)(13)(14)(15)(16)(19)(20)(21). Both ␣v␤3 and ␣v␤6 integrins have been proposed as cell membrane receptors for FMDV in vivo, and we have recently demonstrated, with bovine epithelium, that ␣v␤6 is expressed at considerably higher levels than ␣v␤3 on epithelial cells. The highest levels of ␣v␤6 were detected in epithelial cells of the tongue and interdigital skin, which are sites most likely to show lesion formation in FMDV infection of cattle (P. Monaghan et al., unpublished data).Previous studies of lesion formation have identified areas of increa...

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The ultrastructure of the developing replication site in foot-and-mouth disease virus-infected BHK-38 cells

Cited by 78 publications

References 26 publications

Amino Acid Changes in Proteins 2B and 3A Mediate Rhinovirus Type 39 Growth in Mouse Cells

Amino Acid Changes in Proteins 2B and 3A Mediate Rhinovirus Type 39 Growth in Mouse Cells

Influenza A virus ribonucleoproteins modulate host recycling by competing with Rab11 effectors

Use of Confocal Immunofluorescence Microscopy To Localize Viral Nonstructural Proteins and Potential Sites of Replication in Pigs Experimentally Infected with Foot-and-Mouth Disease Virus

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