RNA synthesis of vesicular stomatitis virus-infected cells: in vivo regulation of replication.
Pulse-labeling of vesicular stomatitis virus-infected HeLa and BHK cells with [3H]uridine throughout the infectious cycle demonstrated two peaks of uridine incorporation into virus-specific RNA molecules. By separating total RNA synthesis into replication and transcription products, we showed that replication occurs over a shorter period of time in one peak synthesis. The biphasic nature of uridine incorporation is in part due to a general membrane phenomenon of reduced metabolite transport during vesicular stomatis virus infection and in part due to the apparent uncoupling of replication and transcription. A change in the ratio of newly synthesized plus and minus strands of the genome length (42S) RNA was found as the infection proceeded. Early in the infection, plus-stranded 42S RNA comprised 40% of the total genome length RNA synthesis, whereas late in infection, only 15 to 20% of the 42S RNA synthesized was complementary to the virion minus strand. Our data suggest that the rate of synthesis of plus-stranded 42S RNA was constant throughout the infection. The rate of virus release was determined by monitoring the uptake of [3H]uridine into released virus particles. Virus maturation and release are closely associated with the assembly of 42S RNA-containing nucleocapsids.
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D006367:HeLa Cells; D006801:Humans; D007668:Kidney; D012367:RNA, Viral / Q000096:biosynthesis*; D014158:Transcription, Genetic; D014529:Uridine / Q000378:metabolism; D014721:Vesicular stomatitis Indiana virus / Q000254:growth & development / Q000378:metabolism*; D014779:Virus Replication
D012367:RNA, Viral; D014529:Uridine