Factors regulating template switch in vitro by viral RNA-dependent RNA polymerases: implications for RNA-RNA recombination.
Copy-choice RNA recombination occurs during viral RNA synthesis when the viral transcription complex switches templates. We demonstrate that RNA-dependent RNA polymerase from bovine viral diarrhea virus and the replicases from three plant-infecting RNA viruses can produce easily detectable recombination products in vitro by switching templates during elongative RNA synthesis. Template sequence and/or structure, and NTP availability affected the frequency of template switch by the transcription complex. Our results provide biochemical support for copy-choice recombination and establish assays for mechanistic analyses of intermolecular RNA recombination in vitro.
Kim
M J
MJ
Department of Biology, Indiana University, 1001 East Third Street, Bloomington, IN 47405, USA.
Kao
C
C
D001483:Base Sequence; D017795:Bromovirus / Q000201:enzymology; D017799:Cucumovirus / Q000201:enzymology; D001908:Diarrhea Viruses, Bovine Viral / Q000201:enzymology* / Q000235:genetics*; D019281:Dimerization; D009690:Nucleic Acid Conformation; D012367:RNA, Viral / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics*; D013698:Templates, Genetic; D014158:Transcription, Genetic
D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase