title: The poliovirus 2C cis-acting replication element-mediated uridylylation of VPg is not required for synthesis of negative-sense genomes.
abstract: Nucleotides in the terminal loop of the poliovirus 2C cis-acting replication element (2C(CRE)), a 61 nt structured RNA, function as the template for the addition of two uridylate (U) residues to the viral protein VPg. This uridylylation reaction leads to the formation of VPgpUpU, which is used by the viral RNA polymerase as a nucleotide-peptide primer for genome replication. Although VPg primes both positive- and negative-strand replication, the specific requirement for 2C(CRE)-mediated uridylylation for one or both events has not been demonstrated. We have used a cell-free in vitro translation and replication reaction to demonstrate that 2C(CRE) is not required for the initiation of the negative-sense strand, which is synthesized in the absence of 2C(CRE)-mediated VPgpUpU formation. We propose that the 3' poly(A) tail could serve as the template for the formation of a VPg-poly(U) primer that functions in the initiation of negative-sense strands.
authors.affiliation: Division of Virology, Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 5JR, UK.
authors.forename: Ian G
authors.initials: IG
authors.lastname: Goodfellow
authors.affiliation: University of California San Francisco, Genentech Hall, San Francisco, CA 94143-2280, USA.
authors.forename: Charlotta
authors.initials: C
authors.lastname: Polacek
authors.affiliation: University of California San Francisco, Genentech Hall, San Francisco, CA 94143-2280, USA.
authors.forename: Raul
authors.initials: R
authors.lastname: Andino
authors.affiliation: Division of Virology, Faculty of Biomedical and Life Sciences, University of Glasgow, Glasgow G11 5JR, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Evans
mesh_terms: D016679:Genome, Viral; D017955:Poliovirus / Q000235:genetics* / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis; D012093:Replicon / Q000502:physiology*; D020218:Response Elements; D014542:Uridine Monophosphate / Q000378:metabolism*; D014758:Viral Core Proteins / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
chemical_list: D012367:RNA, Viral; D014758:Viral Core Proteins; D014542:Uridine Monophosphate