Host Factors Associated with the Sindbis Virus RNA-Dependent RNA Polymerase: Role for G3BP1 and G3BP2 in Virus Replication
Host Factors Associated with the Sindbis Virus RNA-Dependent RNA Polymerase: Role for G3BP1 and G3BP2 in Virus Replication
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DOI：
10.1128/jvi.01983-09
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发表时间：
2010-07-01
影响因子：
5.4
通讯作者：
MacDonald, Margaret R.
中科院分区：
文献类型：
--
作者：
Cristea, Ileana M.;Rozjabek, Heather;MacDonald, Margaret R.
辛德毕斯病毒（SINV）是甲病毒属的原型成员，其成员引起严重的人类疾病，对此没有特异性治疗。为了确定在SINV RNA基因组复制中重要的宿主因素，我们产生了表达nsP 4的SINV，nsP 4是病毒RNA依赖性RNA聚合酶，具有框内3x Flag表位标签。蛋白质组学分析的nsP 4-含有复合物分离的细胞感染的标记病毒揭示了29个相关的宿主蛋白。其中，10种蛋白质仅在感染后期（12小时）相关，14种蛋白质在早期和晚期相关，5种蛋白质仅在早期（感染后6小时）分离。这些结果证明了在感染过程中发生的病毒-宿主相互作用的动态性质，并表明nsP 4执行的多种功能可能需要不同的宿主蛋白。在两次感染中发现的与nsP 4相关的两种相关蛋白，GTP酶激活蛋白（SH 3结构域）结合蛋白1（G3 BP 1）和G3 BP 2先前也被鉴定为与SINV nsP 2和nsP 3相关。我们证明了一个可能重叠的作用，这些主机的因素限制SINV复制事件。本研究还确定了10个主机因素与nsP 4感染后6小时，没有发现与nsP 2或nsP 3。这些因子是在RNA复制过程中发挥重要作用的候选者。确定复制所必需的宿主因素应导致中断甲病毒复制的新策略。
Sindbis virus (SINV) is the prototype member of the Alphavirus genus, whose members cause severe human diseases for which there is no specific treatment. To ascertain host factors important in the replication of the SINV RNA genome, we generated a SINV expressing nsP4, the viral RNA-dependent RNA polymerase, with an in-frame 3 x Flag epitope tag. Proteomic analysis of nsP4-containing complexes isolated from cells infected with the tagged virus revealed 29 associated host proteins. Of these, 10 proteins were associated only at a later time of infection (12 h), 14 were associated both early and late, and five were isolated only at the earlier time (6 h postinfection). These results demonstrate the dynamic nature of the virus-host interaction that occurs over the course of infection and suggest that different host proteins may be required for the multiple functions carried out by nsP4. Two related proteins found in association with nsP4 at both times of infection, GTPase-activating protein (SH3 domain) binding protein 1 (G3BP1) and G3BP2 were also previously identified as associated with SINV nsP2 and nsP3. We demonstrate a likely overlapping role for these host factors in limiting SINV replication events. The present study also identifies 10 host factors associated with nsP4 6 h after infection that were not found to be associated with nsP2 or nsP3. These factors are candidates for playing important roles in the RNA replication process. Identifying host factors essential for replication should lead to new strategies to interrupt alphavirus replication.