Respiratory Syncytial Virus Induces Host RNA Stress Granules To Facilitate Viral Replication
Respiratory Syncytial Virus Induces Host RNA Stress Granules To Facilitate Viral Replication
复制标题
DOI：
10.1128/jvi.00260-10
复制
发表时间：
2010-12-01
影响因子：
5.4
通讯作者：
Crowe, James E., Jr.
中科院分区：
文献类型：
--
作者：
Lindquist, Michael E.;Lifland, Aaron W.;Crowe, James E., Jr.
哺乳动物细胞质RNA应激颗粒在各种应激条件下被诱导，并与宿主mRNA翻译的调节密切相关。几种病毒在感染过程中诱导应激颗粒，但这些结构在病毒复制过程中的确切功能尚不清楚。在本研究中，我们发现呼吸道合胞病毒（RSV）在感染过程中在上皮细胞中诱导宿主应激颗粒。我们还发现，胁迫颗粒与细胞质病毒包涵体不同，通常在胁迫颗粒中发现的RNA结合蛋白HuR在感染过程中也定位于病毒包涵体。有趣的是，我们证明含有应激颗粒的感染细胞比不形成包涵体的感染细胞含有更多的RSV蛋白。为了研究应激颗粒形成在RSV感染中的作用，我们构建了一个稳定的上皮细胞系，其Ras-GAP SH3结构域结合蛋白（G3BP）的表达降低，显示出抑制应激颗粒反应。令人惊讶的是，与G3BP完整表达的细胞相比，RSV在这些细胞中的复制受损。相反，通过RNA干扰敲低HuR并不影响应激颗粒的形成或RSV的复制。最后，使用RSV基因组RNA特异性RNA探针，我们发现病毒RNA主要定位于病毒包涵体，但在感染期间也有一小部分与应激颗粒相互作用。这些结果表明，RSV诱导宿主应激颗粒反应，并优先在已承诺应激反应的宿主细胞中复制。
Mammalian cell cytoplasmic RNA stress granules are induced during various conditions of stress and are strongly associated with regulation of host mRNA translation. Several viruses induce stress granules during the course of infection, but the exact function of these structures during virus replication is not well understood. In this study, we showed that respiratory syncytial virus (RSV) induced host stress granules in epithelial cells during the course of infection. We also showed that stress granules are distinct from cytoplasmic viral inclusion bodies and that the RNA binding protein HuR, normally found in stress granules, also localized to viral inclusion bodies during infection. Interestingly, we demonstrated that infected cells containing stress granules also contained more RSV protein than infected cells that did not form inclusion bodies. To address the role of stress granule formation in RSV infection, we generated a stable epithelial cell line with reduced expression of the Ras-GAP SH3 domain-binding protein (G3BP) that displayed an inhibited stress granule response. Surprisingly, RSV replication was impaired in these cells compared to its replication in cells with intact G3BP expression. In contrast, knockdown of HuR by RNA interference did not affect stress granule formation or RSV replication. Finally, using RNA probes specific for RSV genomic RNA, we found that viral RNA predominantly localized to viral inclusion bodies but a small percentage also interacted with stress granules during infection. These results suggest that RSV induces a host stress granule response and preferentially replicates in host cells that have committed to a stress response.