RNA-protein interactions: Involvement of NS3, NS5, and 3' noncoding regions of Japanese encephalitis virus genomic RNA
RNA-protein interactions: Involvement of NS3, NS5, and 3' noncoding regions of Japanese encephalitis virus genomic RNA
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DOI：
10.1128/jvi.71.5.3466-3473.1997
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发表时间：
1997-05-01
影响因子：
5.4
通讯作者：
Lin, JH
中科院分区：
文献类型：
--
作者：
Chen, CJ;Kuo, MD;Lin, JH
黄病毒日本脑炎病毒(JEV)的复制机制尚不清楚，病毒基因组3‘端的结构在不同的黄病毒中高度保守，提示它们可能作为RNA复制的顺式作用信号，因此可能与细胞或病毒蛋白特异结合。JEV感染细胞提取液中的两种蛋白p71和p110与正链3‘NCR特异结合，并与细胞提取液中JEV RNA的合成水平相关。紫外光交联、免疫印迹和免疫沉淀分析表明，p110和p71蛋白分别为JEV NS5和NS3，它们被认为是RNA复制酶的组成部分，这两种蛋白的结合需要正链和3’NCR中的茎环结构。两种蛋白质在体内可相互作用形成蛋白质-蛋白质复合体，提示JEV基因组RNA的3‘端NCR可能与NS3、NS5形成复制复合体；该复合体可能参与乙脑病毒负链RNA的合成。
The mechanism of replication of the flavivirus Japanese encephalitis virus (JEV) is not well known, The structures at the 3' end of the viral genome are highly conserved among divergent flaviviruses, suggesting that they may function as cis-acting signals for RNA replication and, as such, might specifically bind to cellular or viral proteins, UV cross-linking experiments were performed to identify the proteins that bind with the JEV plus-strand 3' noncoding region (NCR). Two proteins, p71 and p110, from JEV-infected but not from uninfected cell extracts were shown to bind specifically to the plus-strand 3' NCR, The quantities of these binding proteins increased during the course of JEV infection and correlated with the levels of JEV RNA synthesis in cell extracts, UV cross-linking coupled,vith Western blot and immunoprecipitation analysis showed that the p110 and p71 proteins were JEV NS5 and NS3, respectively, which are proposed as components of the RNA replicase, The putative stem-loop structure present within the plus-strand 3' NCR was required for the binding of these proteins, Furthermore, both proteins could interact with each other and form a protein-protein complex in vivo, These findings suggest that the 3' NCR of JEV genomic RNA may form a replication complex together with NS3 and NS5; this complex may be involved in JEV minus-strand RNA synthesis.