Local structural changes of the influenza A virus ribonucleoprotein complex by single mutations in the specific residues involved in efficient genome packaging.
Local structural changes of the influenza A virus ribonucleoprotein complex by single mutations in the specific residues involved in efficient genome packaging.
复制标题
甲型流感病毒核糖核蛋白复合物的局部结构变化是由参与有效基因组包装的特定残基的单突变引起的。
DOI：
10.1016/j.virol.2019.03.004
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发表时间：
2019
期刊：
影响因子：
3.7
通讯作者：
Kurokawa K
中科院分区：
文献类型：
--
作者：
Takizawa N;Ogura Y;Fujita Y;Noda T;Shigematsu H;Hayashi T;Kurokawa K
甲型流感病毒基因组由八个单链负义RNA片段组成。位于每个片段的3′和5′端的非编码区是基因组包装所必需的，而末端编码区则需要精确地捆绑八个片段。然而，对基因组捆绑重要的核苷酸残基没有被定义。在此，我们在血凝素（HA）或基质蛋白2（M2）基因中引入提前终止密码子，并构建了在末端编码区含有随机序列的病毒文库。使用这些病毒库，我们确定了参与有效的病毒繁殖的核苷酸残基。病毒基因组包装受损的病毒，在这些确定的残基含有单一突变。此外，这些单一突变改变了病毒核糖核蛋白复合物的局部结构。我们的研究结果表明，病毒蛋白编码区的特定核苷酸残基参与形成病毒核糖核蛋白复合物的精确结构。
The influenza A virus genome consists of eight single-stranded negative-sense RNA segments. The noncoding regions located at the 3′- and 5′- ends of each segment are necessary for genome packaging, and the terminal coding regions are required to precisely bundle the eight segments. However, the nucleotide residues important for genome bundling are not defined. Here, we introduced premature termination codons in the hemagglutinin (HA) or matrix protein 2 (M2) gene and constructed virus libraries containing random sequences in the terminal coding regions. Using these virus libraries, we identified nucleotide residues involved in efficient virus propagation. Viral genome packaging was impaired in viruses that contained single mutations at these identified residues. Furthermore, these single mutations altered the local structure of the viral ribonucleoprotein complex. Our results show that specific nucleotide residues in the viral protein coding region are involved in forming the precise structure of the viral ribonucleoprotein complex.