Semliki forest virus-specific RNAs synthesized in vitro by enzyme from infected BHK cells.
When Semliki Forest virus (SFV)-infected BHK cells were disrupted 4 h after infection, 75 to 90% of the total virus-specific RNA synthesizing enzyme was found in the large particle fraction, along with 75 to 90% of the in vivo-synthesized double-stranded RNAs. The RNA products of this enzyme-template complex in an in vitro system were double-stranded RNAs sedimenting predominantly at 18S, and single-stranded RNAs sedimenting at 42S, 26S, and 22S. The various virus-specific SFV RNAs synthesized in vitro were associated with different sized structures, and thus each was separable by differential centrifugation. Kinetic and pulse-chase experiments showed that the double-stranded RNAs were the precursors to the single-stranded RNAs. There were several double-stranded RNAs identified both in the in vitro product and also in extracts from infected cells. The major replicative form had a molecular weight of 4.4 x 10(6).
D000818:Animals; D002458:Cell Fractionation; D002460:Cell Line; D002474:Cell-Free System; D002478:Cells, Cultured; D002499:Centrifugation, Density Gradient; D006224:Cricetinae; D012321:DNA-Directed RNA Polymerases / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007668:Kidney; D008854:Microscopy, Electron; D008970:Molecular Weight; D010759:Phosphorus Isotopes; D012367:RNA, Viral / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D012672:Semliki forest virus / Q000032:analysis / Q000254:growth & development / Q000378:metabolism*; D013053:Spectrophotometry; D013698:Templates, Genetic
D010759:Phosphorus Isotopes; D012367:RNA, Viral; D012321:DNA-Directed RNA Polymerases