The herpes simplex virus vhs protein induces endoribonucleolytic cleavage of target RNAs in cell extracts
The herpes simplex virus vhs protein induces endoribonucleolytic cleavage of target RNAs in cell extracts
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DOI：
10.1128/jvi.73.9.7153-7164.1999
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发表时间：
1999-09-01
影响因子：
5.4
通讯作者：
Smiley, JR
中科院分区：
文献类型：
--
作者：
Elgadi, MM;Hayes, CE;Smiley, JR
单纯疱疹病毒病毒体宿主关闭 (vhs) 蛋白（UL41 基因产物）是 HSV 病毒体外皮的一个组成部分，可在 Hm 感染的早期阶段触发宿主蛋白合成的关闭并加速 mRNA 的降解。先前的研究表明，HSV 感染细胞的提取物和部分纯化的 HSV 病毒颗粒显示出 vhs 依赖性 RNase 活性，并且当 vhs 在源自兔网织红细胞的体外翻译系统中表达为唯一的 HSV 蛋白时，足以触发加速 RNA 降解。我们使用兔网织红细胞翻译系统更详细地描述了 vhs 诱导的 RNA 衰减模式。我们在此报道，vhs 依赖性 RNA 衰变通过核糖核酸内切裂解进行，不受 RNA 底物中 5' 帽或 3' 聚 (A) 尾存在的影响，需要 Mg2+，并且在不存在核糖体的情况下发生。有趣的是，优先初始切割位点聚集在一个已详细表征的 RNA 底物的 5' 象限上。伪狂犬病病毒的 vhs 同源物也在该体外系统中诱导加速 RNA 衰减。
The herpes simplex virus virion host shutoff (vhs) protein (UL41 gene product) is a component of the HSV virion tegument that triggers shutoff of host protein synthesis and accelerated mRNA degradation during the early stages of Hm infection. Previous studies have demonstrated that extracts from HSV-infected cells and partially purified HSV virions display vhs-dependent RNase activity and that vhs is sufficient to trigger accelerated RNA degradation when expressed as the only HSV protein in an in vitro translation system derived from rabbit reticulocytes. We have used the rabbit reticulocyte translation system to characterize the mode of vhs-induced RNA decay in more detail. We report here that vhs-dependent RNA decay proceeds through endoribonucleolytic cleavage, is not affected by the presence of a 5' cap or a 3' poly(A) tail in the RNA substrate, requires Mg2+, and occurs in the absence of ribosomes. Intriguingly, sites of preferential initial cleavage were clustered over the 5' quadrant of one RNA substrate that was characterized in detail. The vhs homologue of pseudorabies virus also induced accelerated RNA decay in this in vitro system.