Analysis of herpes simplex virus-induced mRNA destabilizing activity using an in vitro mRNA decay system.
Analysis of herpes simplex virus-induced mRNA destabilizing activity using an in vitro mRNA decay system.
复制标题
使用体外 mRNA 衰减系统分析单纯疱疹病毒诱导的 mRNA 不稳定活性。
DOI：
10.1093/nar/19.16.4459
复制
发表时间：
1991
影响因子：
14.9
通讯作者：
Ross,J
中科院分区：
文献类型：
--
作者：
Sorenson,CM;Hart,PA;Ross,J
大多数宿主mRNA在细胞感染1型单纯疱疹病毒（HSV-1）后不久被降解。这种早期关闭或早期不稳定反应是由病毒体组分，病毒体宿主关闭（vhs）蛋白诱导的。HSV-1突变体vhs 1和vhs-ΔSma产生缺陷或失活的vhs蛋白，不能诱导早期关闭。我们用一个体外mRNA衰变系统来分析去稳定化过程。将来自未感染的人红白血病细胞的多聚体（用作靶mRNA 5的来源）与来自HSV-1或模拟感染的鼠红白血病细胞的多聚体或多聚体后上清液（S130）混合。正常稳定的γ-珠蛋白mRNA被来自野生型病毒感染细胞的S130不稳定约15倍，但不被来自模拟感染细胞或来自用两种HSV突变体中的任一种感染的细胞的S130不稳定。病毒诱导的去稳定活性对两种正常不稳定的mRNA组蛋白和c-myc的体外半衰期没有显著影响。在感染细胞的多聚核糖体中未检测到去稳定活性。我们的结论是，病毒诱导的去稳定剂活性可以在体外发挥作用，位于感染细胞的S130，并加速一些，但不是所有，多核糖体相关的宿主mRNA的衰变率。
Most host mRNAs are degraded soon after infection of cells with herpes simplex virus type 1 (HSV-1). This early shutoff or early destabilization response is induced by a virion component, the virion host shutoff (vhs) protein. HSV-1 mutants,vhs1 andvhs-ΔSma, which produce defective or inactivevhsprotein, fail to induce early shutoff. We have used anin vitromRNA decay system to analyze the destabilization process. Polysomes from uninfected human erythroleukemia cells, used as a source of target mRNA5, were mixed with polysomes or with post-polysomal supernatant (S130) from HSV-1- or mock-infected murine erythroleukemia cells. Normally stable γ-globin mRNA was destabilized by approximately 15-fold with S130 from wild-type virus-infected cells but was not destabilized with S130 from mock-infected cells or from cells infected with either of the two HSV mutants. The virus-induced destabilizing activity had no significant effect on thein vitrohalf-lives of two normally unstable mRNAs, histone and c-myc. No destabilizing activity was detected in polysomes from infected cells. We conclude that a virus-induced destabilizer activity can functionin vitro, is located in the S130 of infected cells, and accelerates the decay rates of some, but not all, polysome-associated host mRNAs.