Distinct Roles of Cellular ESCRT-I and ESCRT-III Proteins in Efficient Entry and Egress of Budded Virions of Autographa californica Multiple Nucleopolyhedrovirus
Distinct Roles of Cellular ESCRT-I and ESCRT-III Proteins in Efficient Entry and Egress of Budded Virions of Autographa californica Multiple Nucleopolyhedrovirus
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细胞 ESCRT-I 和 ESCRT-III 蛋白在苜蓿银纹夜蛾多重核多角体病毒出芽病毒粒子有效进入和排出中的不同作用
DOI：
10.1128/jvi.01636-17
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发表时间：
2017-10
影响因子：
5.4
通讯作者：
Li Zhaofei
中科院分区：
文献类型：
--
作者：
Yue Qi;Yu Qianlong;Yang Qi;Xu Ye;Guo Ya;Blissard Gary;Li Zhaofei
摘要运输所需的内体分选复合体(ESCRT)机制是许多包膜病毒萌发所必需的。最近，研究表明，Vps4是ESCRT-III复合体循环的关键调控因子，是有效感染加州拟银纹夜蛾多核型多角体病毒(AcMNPV)所必需的。然而，ESCRT的组装、调控和功能是复杂的，关于特定ESCRT复合体参与AcMNPV感染的细节知之甚少。在本研究中，从果蝇中克隆了ESCRT-I(Tsg101和Vps28)和ESCRT-III(Vps2B、Vps20、Vps24、Snf7、Vps46和Vps60)的核心成分。利用病毒互补系统和RNA干扰(RNAi)分析，我们发现ESCRT-I和ESCRT-III复合体是AcMNPV有效进入昆虫细胞所必需的。在击倒或过度表达ESCRT-I和ESCRT-III复合体的显性阴性(DN)形式的细胞中，进入细胞的病毒粒子部分被捕获在细胞质中。为了只检查出口，将针对单个ESCRT-I或ESCRT-III基因的双链RNA(DsRNA)以及表达DN型ESCRT-I和ESCRT-III组件的病毒杆菌DNA或病毒杆菌DNA导入细胞。我们发现ESCRT-III组分(而不是ESCRT-I组分)是后代核衣壳有效核出口所必需的。此外，我们还发现一些杆状病毒核心蛋白或保守蛋白(Ac11、Ac76、AC78、GP41、Ac93、Ac103、Ac142和Ac146)与Vps4和ESCRT-III的组成部分相互作用。我们认为这些病毒蛋白可能形成一个“出口复合体”，参与将ESCRT-III组分招募到核膜上的病毒出口区域。重要性ESCRT系统被许多包膜病毒劫持，以调节萌发和释放。最近发现，作为细胞ESCRT机制的关键调节因子，Vps4对于银纹夜蛾多核型多角体病毒(AcMNPV)的有效进出是必需的。然而，关于特异性ESCRT复合体在AcMNPV感染中的作用还知之甚少。在这项研究中，我们证明了ESCRT-I和ESCRT-III复合体是AcMNPV有效进入昆虫细胞所必需的。ESCRT-III的成分(但不是ESCRT-I)对于后代核衣壳的有效核出口也是必要的。一些杆状病毒核心或保守蛋白被发现与Vps4和ESCRT-III的组成部分相互作用，这些相互作用可能表明在病毒核衣壳的核释放或运输过程中形成了“出口复合体”。
ABSTRACT The endosomal sorting complex required for transport (ESCRT) machinery is necessary for budding of many enveloped viruses. Recently, it was demonstrated that Vps4, the key regulator for recycling of the ESCRT-III complex, is required for efficient infection by the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). However, ESCRT assembly, regulation, and function are complex, and little is known regarding the details of participation of specific ESCRT complexes in AcMNPV infection. In this study, the core components of ESCRT-I (Tsg101 and Vps28) and ESCRT-III (Vps2B, Vps20, Vps24, Snf7, Vps46, and Vps60) were cloned from Spodoptera frugiperda. Using a viral complementation system and RNA interference (RNAi) assays, we found that ESCRT-I and ESCRT-III complexes are required for efficient entry of AcMNPV into insect cells. In cells knocking down or overexpressing dominant negative (DN) forms of the components of ESCRT-I and ESCRT-III complexes, entering virions were partially trapped within the cytosol. To examine only egress, cells were transfected with the double-stranded RNA (dsRNA) targeting an individual ESCRT-I or ESCRT-III gene and viral bacmid DNA or viral bacmid DNA that expressed DN forms of ESCRT-I and ESCRT-III components. We found that ESCRT-III components (but not ESCRT-I components) are required for efficient nuclear egress of progeny nucleocapsids. In addition, we found that several baculovirus core or conserved proteins (Ac11, Ac76, Ac78, GP41, Ac93, Ac103, Ac142, and Ac146) interact with Vps4 and components of ESCRT-III. We propose that these viral proteins may form an “egress complex” that is involved in recruiting ESCRT-III components to a virus egress domain on the nuclear membrane. IMPORTANCE The ESCRT system is hijacked by many enveloped viruses to mediate budding and release. Recently, it was found that Vps4, the key regulator of the cellular ESCRT machinery, is necessary for efficient entry and egress of Autographa californica multiple nucleopolyhedrovirus (AcMNPV). However, little is known about the roles of specific ESCRT complexes in AcMNPV infection. In this study, we demonstrated that ESCRT-I and ESCRT-III complexes are required for efficient entry of AcMNPV into insect cells. The components of ESCRT-III (but not ESCRT-I) are also necessary for efficient nuclear egress of progeny nucleocapsids. Several baculovirus core or conserved proteins were found to interact with Vps4 and components of ESCRT-III, and these interactions may suggest the formation of an “egress complex” involved in the nuclear release or transport of viral nucleocapsids.
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影响因子：
64.8
作者：
Feng Z;Hensley L;McKnight KL;Hu F;Madden V;Ping L;Jeong SH;Walker C;Lanford RE;Lemon SM
通讯作者：
Lemon SM
DOI：
10.1083/jcb.201001162
发表时间：
2010-07-26
期刊：
The Journal of cell biology
影响因子：
--
作者：
Ohkawa T;Volkman LE;Welch MD
通讯作者：
Welch MD
艾滋病毒组装部位的ESCRT机械的分布揭示了ESCRT亚基的病毒脚手架。
DOI：
10.1126/science.1247786
发表时间：
2014-02-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Van Engelenburg SB;Shtengel G;Sengupta P;Waki K;Jarnik M;Ablan SD;Freed EO;Hess HF;Lippincott-Schwartz J
通讯作者：
Lippincott-Schwartz J
苜蓿银纹夜蛾核多角体病毒ac93核心基因的鉴定及其核内微泡形成和核衣壳出核的要求
影响因子：
5.4
作者：
Yuan, Meijin;Huang, Zhenqiu;Wei, Denghui;Hu, Zhaoyang;Yang, Kai;Pang, Yi
通讯作者：
Pang, Yi
影响因子：
16.8
作者：
Yang, Bei;Stjepanovic, Goran;Shen, Qingtao;Martin, Andreas;Hurley, James H.
通讯作者：
Hurley, James H.