The specificity of rabies virus RNA encapsidation by nucleoprotein
The specificity of rabies virus RNA encapsidation by nucleoprotein
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DOI：
10.1006/viro.1997.9022
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发表时间：
1998-03-01
期刊：
影响因子：
3.7
通讯作者：
Fu, ZF
中科院分区：
文献类型：
--
作者：
Yang, J;Hooper, DC;Fu, ZF
狂犬病病毒核蛋白（N）在体内与负链基因组RNA结合，该RNA-N复合物与病毒磷蛋白（P）和RNA聚合酶（L）一起在病毒感染细胞中形成活性胞质核糖核蛋白（RNP）复合物，并在病毒颗粒中形成RNP核心。RNP复合物能够在体内和体外启动病毒RNA转录和复制。为了深入了解导致RNA-N复合物形成的事件，我们研究了狂犬病病毒N和正链前导RNA转录本之间的相互作用。结合研究表明，重组N优先结合狂犬病病毒的前导RNA和N结合的前导RNA是5至10倍强于nonleader RNA。未标记的前导RNA可竞争性抑制N对前导RNA的包封，但非前导RNA则不能抑制N对前导RNA的包封。此外，当P同时加入到N蛋白的双核苷酸化反应中时，非前导RNA的N蛋白双核苷酸化被抑制，而前导RNA的N蛋白双核苷酸化不被抑制，表明P有助于赋予前导RNA被N蛋白双核苷酸化的特异性。前导RNA被N取代的起始信号已经被定位到富含A的RNA序列的核苷酸20-30。用N-缺失突变体的研究表明，完整的N是使RNA糖苷化所必需的，因为N的N-或C-末端的氨基酸残基的缺失消除了N使前导RNA糖苷化的能力。(C)北京：科学出版社.
Rabies virus nucleoprotein (N) encapsidates negative-strand genomic RNA in vivo, and this RNA-N complex, together with the nominal viral phosphoprotein (P) and RNA polymerase (L), forms the active cytoplasmic ribonucleoprotein (RNP) complex in virus-infected cells and the RNP core in virus particles. The RNP complex is capable of initiating viral RNA transcription and replication in vivo and in vitro. To obtain insight into the events leading to the formation of the RNA-N complex, we have investigated the interaction between rabies virus N and the positive-strand leader RNA transcript. Binding studies revealed that recombinant N binds preferentially to rabies virus leader RNA and that N binding to leader RNA was 5 to 10 times stronger than to nonleader RNA. Encapsidation of leader RNA by N could be competetively inhibited by unlabeled leader RNA but not by nonleader RNA. Furthermore, N protein encapsidation of nonleader RNA but not the leader RNA was inhibited when P was simultaneously added into the encapsidation reaction, indicating that P helps confer the specificity of leader RNA encapsidation by N. The initiation signal for leader RNA encapsidation by N has been mapped to nucleotides 20-30 of the RNA sequence which is A rich. Studies with N-deletion mutants indicate that the intact N is required to encapsidate RNA, since deletion of amino acid residues from either the N- or the C-terminus of N abolishes the ability of N to encapsidate leader RNA. (C) 1998 Academic Press.