Purified recombinant bluetongue virus VP1 exhibits RNA replicase activity
Purified recombinant bluetongue virus VP1 exhibits RNA replicase activity
复制标题
DOI：
10.1128/jvi.78.8.3994-4002.2004
复制
发表时间：
2004-04-01
影响因子：
5.4
通讯作者：
Roy, P
中科院分区：
文献类型：
--
作者：
Boyce, M;Wehrfritz, J;Roy, P
所有已知的双链RNA(DsRNA)病毒的聚合酶蛋白都位于一个复杂的亚病毒核心颗粒中，该核心颗粒负责病毒基因组的转录。对于呼肠孤病毒科的成员来说，这种颗粒允许信使正义RNA合成，同时在真核细胞感染期间将病毒基因组从细胞dsRNA监测系统中隔离出来。蓝舌病毒(BTV)的核心颗粒由主要结构蛋白VP3和VP7以及次要的酶蛋白VP1(聚合酶)、VP4(封闭酶)和VP6(解旋酶)组成。在这份报告中，我们描述了在没有BTV核心其他蛋白质的情况下，VP1从病毒正链RNA模板合成dsRNA的全过程。这种复制酶的活性包括从头合成的开始，然后是负链的延长。纯化的VP1对BTV正链模板几乎没有序列特异性，这表明选择病毒而不是非病毒RNA模板来自于它与病毒核心内其他蛋白质的结合。
The polymerase protein of all known double-stranded RNA (dsRNA) viruses is located within a complex subviral core particle that is responsible for transcription of the viral genome. For members of the family Reoviridae, this particle allows messenger sense RNA synthesis while sequestering the viral genome away from cellular dsRNA surveillance systems during infection of eukaryotic cells. The core particle of bluetongue virus (BTV) consists of the major structural proteins VP3 and VP7 and the minor enzymatic proteins VP1 (polymerase), VP4 (capping enzyme), and VP6 (helicase). In this report we have characterized fully processive dsRNA synthesis by VP1 from a viral plus-strand RNA template in the absence of the other proteins of the BTV core. This replicase activity consists of de novo initiation of synthesis, followed by elongation of the minus strand. Purified VP1 exhibits little sequence specificity for BTV plus-strand template, suggesting that the choice of viral over nonviral RNA template comes from its association with other proteins within the viral core.