Studies on the Infectivity of Foot-and-Mouth Disease Virus RNA using Microinjection

Belsham, Graham J.; Bostock, Christopher J.

doi:10.1099/0022-1317-69-2-265

Cited by 27 publications

(10 citation statements)

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“…Most picornaviruses replicate with high efficiency within susceptible cells, and within a few hours, the amount of viral RNA can represent 5% of the total RNA in cells (a level similar to that of all the cellular cytoplasmic mRNAs together). Nearly all of the FMDV RNA generated by replication is infectious; indeed, microinjection of cells with as little as 1 to 2 molecules of viral RNA is sufficient to initiate an infection (4). Thus, the replication of FMDV genomes within cells is remarkably efficient.…”

mentioning

confidence: 93%

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D ^pol ) In Vitro

Nayak

Goodfellow

Belsham

2005

J Virol

101

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The 5 terminus of picornavirus genomic RNA is covalently linked to the virus-encoded peptide 3B (VPg). Foot-and-mouth disease virus (FMDV) is unique in encoding and using 3 distinct forms of this peptide. These peptides each act as primers for RNA synthesis by the virus-encoded RNA polymerase 3D pol . To act as the primer for positive-strand RNA synthesis, the 3B peptides have to be uridylylated to form VPgpU(pU). For certain picornaviruses, it has been shown that this reaction is achieved by the 3D pol in the presence of the 3CD precursor plus an internal RNA sequence termed a cis-acting replication element (cre). The FMDV cre has been identified previously to be within the 5 untranslated region, whereas all other picornavirus cre structures are within the viral coding region. The requirements for the in vitro uridylylation of each of the FMDV 3B peptides has now been determined, and the role of the FMDV cre (also known as the 3B-uridylylation site, or bus) in this reaction has been analyzed. The poly(A) tail does not act as a significant template for FMDV 3B uridylylation.Picornaviruses, including foot-and-mouth disease virus (FMDV), poliovirus (PV), and human rhinoviruses (HRVs), have a positive-sense RNA genome of about 8 kb that is infectious (1, 5). Following virus attachment and entry into the cell, the RNA genome is delivered into the cytoplasm and translation of the genome is required to produce the viral proteins that are necessary for virus assembly and RNA replication. The genome encodes a single large polyprotein that is processed, largely by internal trans-acting proteases, to produce about 12 mature proteins plus various precursors (some of these have distinct functions). The proteins encoded within the P1 region form the capsid, while proteins encoded in the P2 and P3 regions are required for RNA replication. After some rounds of translation, there has to be a switch so that translation of the viral RNA is stopped and RNA replication can commence, since these two processes appear incompatible on the same RNA molecule (11). Most picornaviruses replicate with high efficiency within susceptible cells, and within a few hours, the amount of viral RNA can represent 5% of the total RNA in cells (a level similar to that of all the cellular cytoplasmic mRNAs together). Nearly all of the FMDV RNA generated by replication is infectious; indeed, microinjection of cells with as little as 1 to 2 molecules of viral RNA is sufficient to initiate an infection (4). Thus, the replication of FMDV genomes within cells is remarkably efficient.To replicate the positive-sense genome, an antisense RNA has to be synthesized which then functions as the template for the production of new positive-sense infectious genomes (32). RNA is synthesized by the viral 3D protein that functions as an RNA-dependent RNA polymerase and will be referred to as 3D pol . Interestingly, 3D pol requires the uridylylated form of the 3B/VPg peptide (VPgpU or VPgpUpU) to act as the primer for both positive-and negative-strand synthesis. In recent...

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confidence: 93%

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D ^pol ) In Vitro

Nayak

Goodfellow

Belsham

2005

J Virol

101

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“…28 nm which contains 60 copies of 4 different structural proteins, 1A (VP4), 1B (VP2), 1C (VP3) and 1D (VP1) (for review, see [3]). The viral RNA is sufficient to initiate replication when introduced into the cytoplasm of cells (e.g., see [4], [5]), without any requirement for viral proteins Thus the capsid serves to protect the RNA when the virus is outside of cells and to facilitate delivery of the genome into the cytoplasm of cells in which replication occurs [3].…”

Section: Introductionmentioning

confidence: 99%

Rescue of Foot-and-Mouth Disease Viruses That Are Pathogenic for Cattle from Preserved Viral RNA Samples

et al. 2011

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BackgroundFoot and mouth disease is an economically important disease of cloven-hoofed animals including cattle, sheep and pigs. It is caused by a picornavirus, foot-and-mouth disease virus (FMDV), which has a positive sense RNA genome which, when introduced into cells, can initiate virus replication.Principal FindingsA system has been developed to rescue infectious FMDV from RNA preparations generated from clinical samples obtained under experimental conditions and then applied to samples collected in the “field”. Clinical samples from suspect cases of foot-and-mouth disease (FMD) were obtained from within Pakistan and Afghanistan. The samples were treated to preserve the RNA and then transported to National Veterinary Institute, Lindholm, Denmark. Following RNA extraction, FMDV RNA was quantified by real-time RT-PCR and samples containing significant levels of FMDV RNA were introduced into susceptible cells using electroporation. Progeny viruses were amplified in primary bovine thyroid cells and characterized using antigen ELISA and also by RT-PCR plus sequencing. FMD viruses of three different serotypes and multiple lineages have been successfully rescued from the RNA samples. Two of the rescued viruses (of serotype O and Asia 1) were inoculated into bull calves under high containment conditions. Acute clinical disease was observed in each case which spread rapidly from the inoculated calves to in-contact animals. Thus the rescued viruses were highly pathogenic. The availability of the rescued viruses enabled serotyping by antigen ELISA and facilitated genome sequencing.ConclusionsThe procedure described here should improve the characterization of FMDVs circulating in countries where the disease is endemic and thus enhance disease control globally.

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“…FMDV RNA is infectious by itself, when transfected into susceptible cells [27]. This feature has made possible the construction of infectious cDNA clones [183,253], and their use is a powerful tool to study different genes and functional motives of the viral RNA by the analysis of derivatives bearing mutations or deletions at preselected genomic sites (see Sects.…”

Section: Genomic Organisationmentioning

confidence: 99%

Foot-and-mouth disease virus: a long known virus, but a current threat

et al. 2001

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-Foot-and-mouth disease virus (FMDV) was the first animal virus identified. Since then, FMDV has become a model system in animal virology and a considerable amount of information on its structure, biology and vaccinology has been obtained. However, the disease that this virus produces (FMD) still constitutes one of the main animal health concerns. In this review, we have attempted to summarise the state of the knowledge in different basic and applied areas of FMDV research, with emphasis on those aspects relevant to the control of the disease. FMDV / structure / immunity / vaccine / variability / diagnosisRésumé -Le virus de la fièvre aphteuse : un virus connu de longue date, qui demeure une menace. Le virus de la fièvre aphteuse a été le premier virus animal identifié. Depuis lors, il est devenu un système modèle en virologie animale, et une quantité importante d'informations sur sa structure, sa biologie et sa vaccinologie a été obtenue. Cependant la maladie provoquée par ce virus constitue encore une inquiétude majeure en santé animale. Dans cette revue, nous avons tenté de résumer l'état des connaissances dans différents domaines de recherche, à la fois fondamentaux et appliqués, sur le virus de la fièvre aphteuse, en mettant l'accent sur les aspects relatifs au contrôle de la maladie. virus de la fièvre aphteuse / immunité / vaccin / variabilité / diagnostic

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Studies on the Infectivity of Foot-and-Mouth Disease Virus RNA using Microinjection

Cited by 27 publications

References 14 publications

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D ^pol ) In Vitro

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D ^pol ) In Vitro

Rescue of Foot-and-Mouth Disease Viruses That Are Pathogenic for Cattle from Preserved Viral RNA Samples

Foot-and-mouth disease virus: a long known virus, but a current threat

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Studies on the Infectivity of Foot-and-Mouth Disease Virus RNA using Microinjection

Cited by 27 publications

References 14 publications

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D pol ) In Vitro

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D pol ) In Vitro

Rescue of Foot-and-Mouth Disease Viruses That Are Pathogenic for Cattle from Preserved Viral RNA Samples

Foot-and-mouth disease virus: a long known virus, but a current threat

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Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D ^pol ) In Vitro

Factors Required for the Uridylylation of the Foot-and-Mouth Disease Virus 3B1, 3B2, and 3B3 Peptides by the RNA-Dependent RNA Polymerase (3D ^pol ) In Vitro