The two-stage interaction of Ebola virus VP40 with nucleoprotein results in a switch from viral RNA synthesis to virion assembly/ budding  

作　　者：Linjuan Wu Dongning Jin Dan Wang Xuping Jing Peng Gong Yali Qin Mingzhou Chen 

机构地区：[1]State Key Laboratory of Virology and Modern Virology Research Center,College of Life Sciences,Wuhan University,Wuhan 430072,China [2]Wuhan Institute of Virology,Chinese Academy of Sciences,Wuhan 430071,China

出　　处：《Protein & Cell》2022年第2期120-140,共21页蛋白质与细胞（英文版）

基　　金：We acknowledge Dr.Bo Zhang(Wuhan Institute of Virology)and Dr.Heinz Feldmann(National Institutes of Health,Hamilton,Montana,USA)for providing the Ebola cDNA and minigenome assay system,respectively;This research is supported by grants from the National Natural Science Foundation of China(81825015);National Key R&D Program of China(2017YFA0505801);National Science and Technology Major Project(2018ZX10101004);National Natural Science Foundation of China(81871650 and 31630086);The Natural Science Foundation of Hubei Province Innovation Group(2017CFA022);Advanced Customer Cultivation Project of Wuhan National Biosafety Laboratory(2019ACCP-MS06).

摘　　要：Ebola virus(EBOV)is an enveloped negative-sense RNA virus and a member of the filovirus family.Nucleoprotein(NP)expression alone leads to the formation of inclusion bodies(IBs),which are critical for viral RNA synthesis.The matrix protein,VP40,not only plays a critical role in virus assembly/budding,but also can regulate transcription and replication of the viral genome.However,the molecular mechanism by which VP40 regulates viral RNA synthesis and virion assembly/budding is unknown.Here,we show that within IBs the N-terminus of NP recruits VP40 and is required for VLP-containing NP release.Furthermore,we find four point mutations(L692A,P697A,P698A and W699A)within the C-terminal hydrophobic core of NP result in a stronger VP40–NP interaction within IBs,sequestering VP40 within IBs,reducing VP40–VLP egress,abolishing the incorporation of NC-like structures into VP40–VLP,and inhibiting viral RNA synthesis,suggesting that the interaction of N-terminus of NP with VP40 induces a conformational change in the C-terminus of NP.Consequently,the C-terminal hydrophobic core of NP is exposed and binds VP40,thereby inhibiting RNA synthesis and initiating virion assembly/budding.

关 键 词：Ebola virus NUCLEOPROTEIN matrix protein two-stage interaction RNA synthesis NUCLEOCAPSID assembly/budding 

分 类 号：R37[医药卫生—病原生物学]