A poliovirus replicon containing the chloramphenicol acetyltransferase gene can be used to study the replication and encapsidation of poliovirus RNA

Percy, N; Barclay, Wendy S.; Sullivan, Michael L.; Almond, Jeffrey W.

doi:10.1128/jvi.66.8.5040-5046.1992

Cited by 66 publications

(56 citation statements)

References 30 publications

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“…The VEE, Kunjin, and Sindbis replicon packaging systems involve the cotransfection of replicon and helper RNAs that express the structural genes (59). In addition to this method, replicon RNAs have been packaged by the expression of structural genes in trans from wild-type or mutated virus (53,56). An important concern with these types of replicon systems is the production of recombinant virus, especially when considering the development of a replicon particle vaccine.…”

Section: Discussionmentioning

confidence: 99%

Heterologous Gene Expression from Transmissible Gastroenteritis Virus Replicon Particles

Curtis¹,

Yount

Baric

2002

J Virol

134

162

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We have recently isolated a transmissible gastroenteritis virus (TGEV) infectious construct designatedTransmissible gastroenteritis (TGE) is an economically important, acute enteric disease of swine, which is often 100% fatal in newborn piglets (23,24,46). TGE virus (TGEV), the causative agent of TGE, is a member of the Coronaviridae family and the order Nidovirales. In addition to the Coronaviridae, the order Nidovirales also includes the Arteriviridae family, of which the swine pathogen porcine reproductive and respiratory syndrome virus is a member (12,18,70). Despite significant size differences (ϳ13 to 32 kb), the polycistronic genome organization and regulation of gene expression from a nested set of subgenomic mRNAs are similar for all members of the order (18, 71).TGEV possesses a single-stranded, positive-sense ϳ28.5-kb RNA genome enclosed in a helical nucleocapsid structure that is surrounded by an envelope containing three viral proteins, including the S glycoprotein, the membrane (M) glycoprotein and a small envelope (E) protein (22,25,60,61). Remarkably, only the E and M proteins are absolutely required for particle formation, defining a novel model for virion budding (27, 77). The TGEV genome contains eight large open reading frames (ORFs), which are expressed from full-length or subgenomic-length mRNAs during infection (22,68,69). The 5Ј-most ϳ20 kb contains the replicase genes in two ORFs, 1A and 1B, the latter of which is expressed by ribosomal frameshifting (2, 22). The 3Ј-most ϳ9 kb of the TGEV genome contains the structural genes, each preceded by a highly conserved intergenic sequence (IS) of 7 to 17 nucleotides (nt) in length that functions in the synthesis of each of the subgenomic RNAs (14,22,25,73). The subgenomic mRNAs are arranged in a coterminal nested set structure from the 3Ј end of the genome, and each contains a leader RNA sequence derived from the 5Ј end of the genome. Although each mRNA is polycistronic, the 5Ј-most ORF is preferentially translated, necessitating the synthesis of a distinct mRNA species for each ORF (45,49,68,69). Both full-length and subgenomic-length negativestrand RNAs are also produced and have been implicated in mRNA synthesis (8,64,66,68,69). Subgenomic RNA synthesis occurs by a method of discontinuous transcription, most likely by transcription attenuation during negative-strand synthesis (8, 64).The coronavirus E and M proteins function in virion assembly and release, which involve the constitutive secretory pathway of infected cells. Coexpression of the E and M proteins

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Section: Discussionmentioning

confidence: 99%

Heterologous Gene Expression from Transmissible Gastroenteritis Virus Replicon Particles

Curtis¹,

Yount

Baric

2002

J Virol

134

162

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“…Barclay and colleagues used chloramphenicol acetyltransferase (CAT) inserted into the P1 domain as a foreign marker gene (103). These replicons carrying foreign genes could be encapsidated in trans by coinfection with either a vaccinia virus expressing the poliovirus capsid proteins (104) or PV (105). The exquisite specificity of encapsidation was revealed when enteroviruses other than PV were used in coinfections: the PV replicons could not be packaged in trans into capsids of bovine enterovirus, C-cluster coxsackievirus A21, B-cluster coxsackieviruses B3 and B4, rhinovirus 14, or enterovirus 70 (102,103).…”

Section: All Searches For Rna Packaging Signals In Enterovirus Genomementioning

confidence: 99%

Picornavirus Morphogenesis

Jiang

Liu

et al. 2014

Microbiol Mol Biol Rev

201

196

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SUMMARY The Picornaviridae represent a large family of small plus-strand RNA viruses that cause a bewildering array of important human and animal diseases. Morphogenesis is the least-understood step in the life cycle of these viruses, and this process is difficult to study because encapsidation is tightly coupled to genome translation and RNA replication. Although the basic steps of assembly have been known for some time, very few details are available about the mechanism and factors that regulate this process. Most of the information available has been derived from studies of enteroviruses, in particular poliovirus, where recent evidence has shown that, surprisingly, the specificity of encapsidation is governed by a viral protein-protein interaction that does not involve an RNA packaging signal. In this review, we make an attempt to summarize what is currently known about the following topics: (i) encapsidation intermediates, (ii) the specificity of encapsidation (iii), viral and cellular factors that are required for encapsidation, (iv) inhibitors of encapsidation, and (v) a model of enterovirus encapsidation. Finally, we compare some features of picornavirus morphogenesis with those of other plus-strand RNA viruses.

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“…Briefly, the assay involves two poliovirus genomes each containing a different deleterious (and non-reverting) modification that prevents the production of viable progeny. One genome, a sub-genomic replicon (12), was replication competent but did not encode capsid proteins. The other contained a well-understood mutation (designated SL3) in a critical cis-acting replication element (CRE), a defined stemloop structure essential for positive-strand replication (13,14).…”

Section: Introductionmentioning

confidence: 99%

Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination

Woodman

Arnold

Cameron

et al. 2016

Nucleic Acids Research

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Genetic recombination in single-strand, positive-sense RNA viruses is a poorly understand mechanism responsible for generating extensive genetic change and novel phenotypes. By moving a critical cis-acting replication element (CRE) from the polyprotein coding region to the 3′ non-coding region we have further developed a cell-based assay (the 3′CRE-REP assay) to yield recombinants throughout the non-structural coding region of poliovirus from dually transfected cells. We have additionally developed a defined biochemical assay in which the only protein present is the poliovirus RNA dependent RNA polymerase (RdRp), which recapitulates the strand transfer events of the recombination process. We have used both assays to investigate the role of the polymerase fidelity and nucleotide turnover rates in recombination. Our results, of both poliovirus intertypic and intratypic recombination in the CRE-REP assay and using a range of polymerase variants in the biochemical assay, demonstrate that RdRp fidelity is a fundamental determinant of recombination frequency. High fidelity polymerases exhibit reduced recombination and low fidelity polymerases exhibit increased recombination in both assays. These studies provide the basis for the analysis of poliovirus recombination throughout the non-structural region of the virus genome and provide a defined biochemical assay to further dissect this important evolutionary process.

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A poliovirus replicon containing the chloramphenicol acetyltransferase gene can be used to study the replication and encapsidation of poliovirus RNA

Cited by 66 publications

References 30 publications

Heterologous Gene Expression from Transmissible Gastroenteritis Virus Replicon Particles

Heterologous Gene Expression from Transmissible Gastroenteritis Virus Replicon Particles

Picornavirus Morphogenesis

Biochemical and genetic analysis of the role of the viral polymerase in enterovirus recombination

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