In vitro synthesis of Japanese encephalitis virus (JEV) RNA: membrane and nuclear fractions of JEV-infected cells possess high levels of virus-specific RNA polymerase activity.
Japanese encephalitis virus (JEV)-specific RNAs (including 42S RNA) were synthesized in subcellular fractions prepared from infected C6/36 cells. This in vitro RNA synthesis essentially required Mg2+ and four ribonucleotides, and it was enhanced by K+. The amounts of RNA synthesized in vitro (in extracts from JEV-infected cells) increased as a function of time after infection. The RNA-synthetic activity in nuclear fractions was the highest among three kinds of subcellular fractions. Our data showed that nonstructural proteins NS3 and NS5 were membrane-associated proteins. In particular, NS3 was found almost exclusively in the nuclear and membrane fractions. Our results suggest that NS5 and NS3 may play specific role(s) in flavivirus RNA replication.
Takegami
T
T
Institute for Tropical Medicine, Kanazawa Medical University, Ishikawa, Japan.
Hotta
S
S
D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004664:Encephalitis Virus, Japanese / Q000378:metabolism*; D007700:Kinetics; D008274:Magnesium / Q000378:metabolism; D008566:Membranes / Q000378:metabolism; D012316:RNA Nucleotidyltransferases / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis*; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D012265:Ribonucleotides / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism; D014764:Viral Proteins / Q000096:biosynthesis; D014779:Virus Replication
D012367:RNA, Viral; D012265:Ribonucleotides; D014764:Viral Proteins; D012316:RNA Nucleotidyltransferases; D012324:RNA-Dependent RNA Polymerase; D008274:Magnesium