REPLICATION STRATEGY OF KUNJIN VIRUS - EVIDENCE FOR RECYCLING ROLE OF REPLICATIVE FORM RNA AS TEMPLATE IN SEMICONSERVATIVE AND ASYMMETRIC REPLICATION
REPLICATION STRATEGY OF KUNJIN VIRUS - EVIDENCE FOR RECYCLING ROLE OF REPLICATIVE FORM RNA AS TEMPLATE IN SEMICONSERVATIVE AND ASYMMETRIC REPLICATION
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DOI：
10.1016/0042-6822(85)90446-5
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发表时间：
1985-01-01
期刊：
影响因子：
3.7
通讯作者：
WESTAWAY, EG
中科院分区：
文献类型：
--
作者：
CHU, PWG;WESTAWAY, EG
在潜伏期后早期（约15小时），从[非洲绿色猴肾Vero]细胞质中仅分离到3种形式的昆津病毒特异性RNA，即，44 S基因组大小的单链RNA、20 S双链复制形式（RF）和20-28 S部分RNA酶抗性（约70%为复制中间体（RI）。RF和RI仅在LiCl分级分离后通过在水性琼脂糖凝胶中电泳来解决。RI不进入尿素-聚丙烯酰胺凝胶。未经处理或经RNase处理的RI和RF变性后，电泳图中仅出现44 S RNA。RNA聚合酶活性在8小时感染后，在体外细胞质提取物的测定，并在24小时达到最大值，唯一的主要标记产品是RF，微量的游离44 S RNA也产生。这些结果，以及在脉冲和脉冲追逐实验中[3 H]尿苷掺入RI、RF和44 S RNA的动力学，形成了一个模型的基础，在该模型中黄病毒RF作为保守性和（主要）不对称复制的再循环模板发挥作用，每个循环仅合成1条新生链。
Only 3 forms of Kunjin virus-specified RNA were isolated from [African green monkey kidney Vero] cytoplasm early after the latent period (about 15 h) viz., 44 S genomic-sized single-stranded RNA, 20 S double-stranded replicative form (RF) and 20-28 S partially RNase-resistant (.apprx. 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 h postinfection was detected by in vitro assays of cytoplasmic extracts and reached a maximum at 24 h, 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 1 nascent strand is synthesized per cycle.