期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2014
卷号：111
期号：42
页码：15208-15213
DOI：10.1073/pnas.1413268111
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceIn this paper, we reveal several insights into how mumps virus (MuV) replicates its RNA genome. The MuV genomic RNA is packaged by the nucleocapsid protein (N), forming a helical structure called the nucleocapsid. The nucleocapsid is the template for RNA synthesis. MuV genomes cannot be copied unless the viral polymerase (vRdRp) can read the sequestered RNA. The MuV phosphoprotein (P) appears to play a central role in this process. In this paper, we provide the first evidence, to our knowledge, of P inducing the nucleocapsid to uncoil. MuV P uses two separate domains to promote viral RNA synthesis. One domain attaches to the nucleocapsid while the other domain relaxes the helical structure to allow vRdRp to easily read the viral genome. Mumps virus (MuV) is a highly contagious pathogen, and despite extensive vaccination campaigns, outbreaks continue to occur worldwide. The virus has a negative-sense, single-stranded RNA genome that is encapsidated by the nucleocapsid protein (N) to form the nucleocapsid (NC). NC serves as the template for both transcription and replication. In this paper we solved an 18-[IMG]f1.gif" ALT="A" BORDER="0">-resolution structure of the authentic MuV NC using cryo-electron microscopy. We also observed the effects of phosphoprotein (P) binding on the MuV NC structure. The N-terminal domain of P (PNTD) has been shown to bind NC and appeared to induce uncoiling of the helical NC. Additionally, we solved a 25-[IMG]f1.gif" ALT="A" BORDER="0">-resolution structure of the authentic MuV NC bound with the C-terminal domain of P (PCTD). The location of the encapsidated viral genomic RNA was defined by modeling crystal structures of homologous negative strand RNA virus Ns in NC. Both the N-terminal and C-terminal domains of MuV P bind NC to participate in access to the genomic RNA by the viral RNA-dependent-RNA polymerase. These results provide critical insights on the structure-function of the MuV NC and the structural alterations that occur through its interactions with P.
