Replication of Tomato bushy stunt virus RNA in a plant in vitro system
Replication of Tomato bushy stunt virus RNA in a plant in vitro system
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DOI：
10.1016/j.virol.2009.05.009
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发表时间：
2009-08-01
期刊：
影响因子：
3.7
通讯作者：
Behrens, Sven-Erik
中科院分区：
文献类型：
--
作者：
Gursinsky, Torsten;Schulz, Beate;Behrens, Sven-Erik
研究 (+)-链 RNA 病毒复制过程的个体决定因素的理想系统是支持病毒蛋白和 RNA 同步合成的无细胞提取物。在这里，我们将基于烟草细胞胞质提取物的翻译/复制系统应用于番茄丛矮病毒 (TBSV) RNA。体外翻译的 TBSV 蛋白 p33 和 p92 形成病毒复制酶，在同一反应中，完成外源 TBSV DI 或全长 RNA 的整个复制周期。突变 TBSV RNA 的测试证实了体外复制反应的模板特异性。互补实验确定了早期发现的 TBSV 宿主因子的重要性。有趣的是，病毒复制酶的形成也在没有同时蛋白质合成的情况下发生，这表明翻译和 RNA 复制在该系统中没有功能上的联系。因此，我们对植物宿主的无细胞提取物进行的研究证实了早期的发现，并为 TBSV RNA 复制过程提供了新的见解。 (C) 2009 Elsevier Inc. 保留所有权利。
An ideal system to investigate individual determinants of the replication process of (+)-strand RNA viruses is a cell-free extract that supports viral protein and RNA synthesis in a synchronized manner. Here, we applied a translation/replication system based on cytoplasmic extracts of Nicotiana tabacum cells to Tomato bushy stunt virus (TBSV) RNA. In vitro translated TBSV proteins p33 and p92 form viral replicase, which, in the same reaction, accomplishes the entire replication cycle on exogenous TBSV DI or full-length RNA. Tests of mutant TBSV RNAs confirmed the template specificity of the in vitro replication reaction. Complementation experiments ascertained the significance of an earlier identified TBSV host factor. Interestingly, formation of the viral replicase occurs also in the absence of concurrent protein synthesis demonstrating that translation and RNA replication are not functionally linked in this system. Our studies with cell-free extracts of a plant host thus confirmed earlier findings and enabled novel insights into the TBSV RNA replication process. (C) 2009 Elsevier Inc. All rights reserved.