title: 'Primer alignment-and-extension': a novel mechanism of viral RNA recombination responsible for the rescue of inactivated poliovirus cDNA clones.
abstract: In the course of experiments designed to assess the potential role of alternative open reading frames (ORF) present in the 5'-terminal untranslated region (5'-UTR) of poliovirus type 1 (Mahoney strain) genomic RNA, we came across a double mutation that completely abrogated the infectivity of full-length cDNA clones. The infectivity was rescued in trans by cotransfecting COS-1 cells with short RNA transcripts of the wild-type 5'-UTR of poliovirus type 2 Lansing, provided a free 3'-OH was available. Direct sequencing of the viral RNA revealed that the infectious viruses recovered were recombinants Lansing/Mahoney, with variable points of 'crossing-over'. A novel mechanism of RNA-RNA recombination, which we propose to call 'primer alignment-and-extension', is described that would explain the high rate of recombination of RNA viruses observed in natural conditions.
authors.affiliation: Department of Cellular and Developmental Biology, University of Rome 'La Sapienza', Viale di Porta Tiburtina 28, 00185-Rome, Italy1.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Pierangeli
authors.affiliation: Department of Cellular and Developmental Biology, University of Rome 'La Sapienza', Viale di Porta Tiburtina 28, 00185-Rome, Italy1.
authors.forename: M
authors.initials: M
authors.lastname: Bucci
authors.affiliation: Department of Cellular and Developmental Biology, University of Rome 'La Sapienza', Viale di Porta Tiburtina 28, 00185-Rome, Italy1.
authors.forename: M
authors.initials: M
authors.lastname: Forzan
authors.affiliation: Department of Cellular and Developmental Biology, University of Rome 'La Sapienza', Viale di Porta Tiburtina 28, 00185-Rome, Italy1.
authors.forename: P
authors.initials: P
authors.lastname: Pagnotti
authors.affiliation: Istituto Superiore di Sanità, 00161-Rome, Italy2.
authors.forename: M
authors.initials: M
authors.lastname: Equestre
authors.affiliation: Department of Cellular and Developmental Biology, University of Rome 'La Sapienza', Viale di Porta Tiburtina 28, 00185-Rome, Italy1.
authors.forename: R
authors.initials: R
authors.lastname: Pérez Bercoff
mesh_terms: D020121:5' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D018387:Codon, Initiator; D018076:DNA, Complementary; D004279:DNA, Viral; D016679:Genome, Viral; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D017955:Poliovirus / Q000235:genetics*; D014176:Protein Biosynthesis; D012313:RNA; D012367:RNA, Viral; D011995:Recombination, Genetic; D014162:Transfection; D014709:Vero Cells; D014771:Virion
chemical_list: D020121:5' Untranslated Regions; D018387:Codon, Initiator; D018076:DNA, Complementary; D004279:DNA, Viral; C034477:RNA primers; D012367:RNA, Viral; D012313:RNA