Small Noncoding RNA Modulates Japanese Encephalitis Virus Replication and Translation in trans
Small Noncoding RNA Modulates Japanese Encephalitis Virus Replication and Translation in trans
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DOI：
10.1186/1743-422x-8-492
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发表时间：
2011-11-01
期刊：
影响因子：
4.8
通讯作者：
Chang, Ruey-Yi
中科院分区：
文献类型：
--
作者：
Fan, Yi-Hsin;Nadar, Muthukumar;Chang, Ruey-Yi
背景：已知日本脑炎病毒 (JEV) 3'-非翻译区 (UTR) 中的序列和结构元件可调节翻译和复制。我们之前报道了小亚基因组黄病毒 RNA (sfRNA) 的丰富积累，其与 JEV 感染细胞中 3'-UTR 的高度保守区域共线。然而，sfRNA 在 JEV 生命周期中的功能仍然未知。 结果：Northern 印迹和实时 RT-PCR 分析表明，sfRNA 在负链 RNA（反基因组）达到平台期的时间点变得明显，表明 sfRNA 在反基因组合成的调节中发挥作用。为了抵消正义sfRNA的影响，将负义sfRNA转染到JEV感染的细胞中，导致反基因组水平升高，表明sfRNA的存在抑制反基因组合成。使用含有与 JEV 部分编码区融合且两侧为相应 JEV UTR 的荧光素酶基因的报告基因 mRNA，研究了 sfRNA 对 JEV 翻译的反式作用效应。体内和体外翻译表明 sfRNA 抑制 JEV 翻译。结论：我们的结果表明 sfRNA 调节病毒的反式翻译和复制。
Background: Sequence and structural elements in the 3'-untranslated region (UTR) of Japanese encephalitis virus (JEV) are known to regulate translation and replication. We previously reported an abundant accumulation of small subgenomic flaviviral RNA (sfRNA) which is collinear with the highly conserved regions of the 3'-UTR in JEV-infected cells. However, function of the sfRNA in JEV life cycle remains unknown.Results: Northern blot and real-time RT-PCR analyses indicated that the sfRNA becomes apparent at the time point at which minus-strand RNA (antigenome) reaches a plateau suggesting a role for sfRNA in the regulation of antigenome synthesis. Transfection of minus-sense sfRNA into JEV-infected cells, in order to counter the effects of plus-sense sfRNA, resulted in higher levels of antigenome suggesting that the presence of the sfRNA inhibits antigenome synthesis. Trans-acting effect of sfRNA on JEV translation was studied using a reporter mRNA containing the luciferase gene fused to partial coding regions of JEV and flanked by the respective JEV UTRs. In vivo and in vitro translation revealed that sfRNA inhibited JEV translation.Conclusions: Our results indicate that sfRNA modulates viral translation and replication in trans.