Chinchar V G, Han J, Mao J, Brooks I, Srivastava K
Department of Microbiology, University of Mississippi Medical Center, Jackson 39216.
Virology. 1994 Aug 15;203(1):187-92. doi: 10.1006/viro.1994.1473.
Cloned DNA restriction fragments encoding representative frog virus 3 messages were used as probes to assess the stability of viral transcripts in infected fathead minnow cells. Analysis of Northern blot hybridization profiles confirmed earlier findings and revealed that in infected cells the steady-state level of representative frog virus 3 (FV3) messages increased throughout the replication cycle. However, when actinomycin D was added at 4 hr after infection to block the synthesis of new transcripts, viral messages were observed to turn over rapidly, with half-lives of approximately 2 hr. These results indicate that viral transcripts were not preferentially stabilized in FV3-infected cells and suggest that the high steady-state level of viral messages present at late times after infection was due to viral transcription outpacing message degradation. Moreover, the instability of viral messages challenges the suggestion that the terminal dyad symmetry (hairpin structure) observed in all frog virus 3 messages sequenced to date plays a role in transcript stability.
编码代表性蛙病毒3信使的克隆DNA限制片段被用作探针，以评估病毒转录本在感染的黑头软口鲦鱼细胞中的稳定性。Northern印迹杂交图谱分析证实了早期的发现，并揭示在感染细胞中，代表性蛙病毒3（FV3）信使的稳态水平在整个复制周期中都有所增加。然而，当在感染后4小时添加放线菌素D以阻断新转录本的合成时，观察到病毒信使迅速周转，半衰期约为2小时。这些结果表明，病毒转录本在FV3感染的细胞中没有优先稳定化，并表明感染后期出现的病毒信使的高稳态水平是由于病毒转录超过了信使降解。此外，病毒信使的不稳定性对以下观点提出了挑战，即迄今为止测序的所有蛙病毒3信使中观察到的末端二联体对称性（发夹结构）在转录本稳定性中起作用。
