State Key Laboratory of Virology, Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Virology. 2010 Dec 20;408(2):138-45. doi: 10.1016/j.virol.2010.09.033. Epub 2010 Oct 20.
We report a genetic interplay among three pairs of long-distance RNA interactions that are involved in West Nile virus (WNV) genome cyclization and replication: 5'CS/3'CSI (conserved sequence), 5'UAR/3'UAR (upstream AUG region), and 5'DAR/3'DAR (downstream AUG region). Deletion of the complete 3'CSI element is lethal for WNV replication, but the replication of the 3'CSI deletion virus could be rescued by second site mutations. Functional analysis, using a genome-length RNA and replicon, mapped the compensatory mutations to the 5'UAR/3'UAR and 5'DAR/3'DAR regions. Biochemical analysis showed that the 3'CSI deletion abolished the 5' and 3' RNA interaction of the genome; the compensatory mutations could partially restore the 5' and 3' genome cyclization. These results demonstrate, for the first time, that a flavivirus without 3'CSI could restore genome cyclization and viral replication through enhancement of the 5'UAR/3'UAR and 5'DAR/3'DAR interactions.
我们报告了三个长距离 RNA 相互作用对之间的遗传相互作用，这些相互作用参与了西尼罗河病毒（WNV）基因组环化和复制：5'CS/3'CSI（保守序列）、5'UAR/3'UAR（上游 AUG 区）和 5'DAR/3'DAR（下游 AUG 区）。完整的 3'CSI 元件缺失对 WNV 复制是致命的，但 3'CSI 缺失病毒的复制可以通过第二点突变来挽救。使用全长 RNA 和复制子的功能分析将补偿突变映射到 5'UAR/3'UAR 和 5'DAR/3'DAR 区域。生化分析表明，3'CSI 缺失消除了基因组的 5'和 3'RNA 相互作用；补偿突变可以部分恢复 5'和 3'基因组环化。这些结果首次表明，没有 3'CSI 的黄病毒可以通过增强 5'UAR/3'UAR 和 5'DAR/3'DAR 相互作用来恢复基因组环化和病毒复制。
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