title: Bluetongue virus VP1 polymerase activity in vitro: template dependency, dinucleotide priming and cap dependency.
abstract: BACKGROUND
  Bluetongue virus (BTV) protein, VP1, is known to possess an intrinsic polymerase function, unlike rotavirus VP1, which requires the capsid protein VP2 for its catalytic activity. However, compared with the polymerases of other members of the Reoviridae family, BTV VP1 has not been characterized in detail.
  
  
  RESULTS
  Using an in vitro polymerase assay system, we demonstrated that BTV VP1 could synthesize the ten dsRNAs simultaneously from BTV core-derived ssRNA templates in a single in vitro reaction as well as genomic dsRNA segments from rotavirus core-derived ssRNA templates that possess no sequence similarity with BTV. In contrast, dsRNAs were not synthesized from non-viral ssRNA templates by VP1, unless they were fused with specific BTV sequences. Further, we showed that synthesis of dsRNAs from capped ssRNA templates was significantly higher than that from uncapped ssRNA templates and the addition of dinucleotides enhanced activity as long as the last base of the dinucleotide complemented the 3' -terminal nucleotide of the ssRNA template.
  
  
  CONCLUSIONS
  We showed that the polymerase activity was stimulated by two different factors: cap structure, likely due to allosteric effect, and dinucleotides due to priming. Our results also suggested the possible presence of cis-acting elements shared by ssRNAs in the members of family Reoviridae.
authors: Faculty of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, London, United Kingdom.
authors: Eiko
authors: E
authors: Matsuo
authors: Polly
authors: P
authors: Roy
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D001483:Base Sequence; D055162:Biocatalysis; D001820:Bluetongue virus / Q000201:enzymology*; D002460:Cell Line; D012315:RNA Caps / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012330:RNA, Double-Stranded / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012367:RNA, Viral / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012324:RNA-Dependent RNA Polymerase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D012315:RNA Caps; D012330:RNA, Double-Stranded; D012367:RNA, Viral; D012324:RNA-Dependent RNA Polymerase