Replication strategy of Kunjin virus: evidence for recycling role of replicative form RNA as template in semiconservative and asymmetric replication.
Only three forms of Kunjin virus-specified RNA were isolated from cytoplasm early after the latent period (about 15 hr) viz., 44 S genomic-sized single-stranded RNA, 20 S double-stranded "replicative form" (RF), and 20-28 S partially ribonuclease-resistant (about 70%) "replicative intermediate" (RI). The RF and RI were resolved by electrophoresis in aqueous-agarose gel only following LiCl fractionation. The RI did not enter urea-polyacrylamide gels. After denaturation of untreated or RNase-treated RI and RF, only 44 S RNA was present in electropherograms. RNA polymerase activity at 8 hr postinfection was detected by in vitro assays of cytoplasmic extracts and reached a maximum at 24 hr, the only major labeled product being RF; a trace amount of free 44 S RNA was also produced. These results, and the kinetics of incorporation of [3H]uridine into RI, RF, and 44 S RNA in pulse and pulse-chase experiments, formed the basis of a model in which flavivirus RF functions as a recycling template for semiconservative and (mainly) asymmetric replication, on which only one nascent strand is synthesized per cycle.
Chu
P W
PW
Westaway
E G
EG
D000818:Animals; D002460:Cell Line; D002472:Cell Transformation, Viral; D002522:Chlorocebus aethiops; D004591:Electrophoresis, Polyacrylamide Gel; D005416:Flavivirus / Q000235:genetics*; D007668:Kidney; D007700:Kinetics; D012313:RNA / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D014779:Virus Replication
D012367:RNA, Viral; D012313:RNA