A unique role for the host ESCRT proteins in replication of Tomato bushy stunt virus.
A unique role for the host ESCRT proteins in replication of Tomato bushy stunt virus.
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DOI：
10.1371/journal.ppat.1000705
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发表时间：
2009-12
期刊：
影响因子：
6.7
通讯作者：
Nagy PD
中科院分区：
文献类型：
--
作者：
Barajas D;Jiang Y;Nagy PD
正链RNA病毒通过组装由病毒和宿主编码蛋白组成的病毒复制酶复合物在感染细胞中复制。先前在酵母模型宿主中对番茄丛状突病毒（TBSV）进行的全基因组筛选显示，7种ESCRT（运输所需的内体分选复合体）蛋白参与了病毒复制。在本文中，我们发现显性阴性Vps23p、Vps24p、Snf7p和Vps4p ESCRT因子的表达抑制了病毒在植物宿主中的复制，这表明tombusvirus通过选择ESCRT蛋白来组装病毒复制酶复合体。我们还发现TBSV p33复制蛋白与Vps23p ESCRT- i和Bro1p辅助ESCRT因子相互作用。与p33的相互作用导致Vps23p被招募到TBSV复制的过氧化物酶体上。当从vps23Δ或vps24Δ酵母中提取时，病毒复制酶的活性降低，复制酶中的负链病毒RNA更容易被核糖核酸酶获取，这表明在没有ESCRT蛋白的情况下，病毒RNA的保护作用在复制酶复合体中受到损害。复制酶复合体的精确组装需要ESCRT蛋白的募集，这可能有助于病毒逃避宿主防御监视系统的识别和/或防止基因沉默机制对病毒RNA的破坏。正链RNA病毒是人类、动物和植物的重要病原体，通过组装由病毒和宿主编码蛋白质组成的病毒复制酶复合物在感染细胞中进行复制。在本文中，我们证明了一组称为ESCRT蛋白（运输所需的内体分选复合物）的宿主因子在tombusvirus复制中起重要作用。ESCRT因子显性阴性突变体的表达抑制了病毒在植物宿主中的复制，这表明tombusvirus通过选择ESCRT蛋白来组装病毒复制酶复合体。此外，我们发现病毒p33复制蛋白与Vps23p ESCRT- i和Bro1p辅助ESCRT因子之间存在直接相互作用。与p33的相互作用导致Vps23p聚集到过氧化物酶体上，这是tombusvirus复制的位点。我们还发现，在没有ESCRT因子的情况下，病毒复制酶复合体内的病毒RNA对核糖核酸酶变得更加敏感，这表明在没有ESCRT蛋白的情况下，病毒RNA的保护作用在复制酶复合体内受到损害。有趣的是，宿主ESCRT因子还影响几种包膜病毒的出芽、细胞内蛋白质转运和细胞质分裂。总的来说，这项工作证明了正链RNA病毒以一种独特的方式使用内体分选途径。
Plus-stranded RNA viruses replicate in infected cells by assembling viral replicase complexes consisting of viral- and host-coded proteins. Previous genome-wide screens with Tomato bushy stunt tombusvirus (TBSV) in a yeast model host revealed the involvement of seven ESCRT (endosomal sorting complexes required for transport) proteins in viral replication. In this paper, we show that the expression of dominant negative Vps23p, Vps24p, Snf7p, and Vps4p ESCRT factors inhibited virus replication in the plant host, suggesting that tombusviruses co-opt selected ESCRT proteins for the assembly of the viral replicase complex. We also show that TBSV p33 replication protein interacts with Vps23p ESCRT-I and Bro1p accessory ESCRT factors. The interaction with p33 leads to the recruitment of Vps23p to the peroxisomes, the sites of TBSV replication. The viral replicase showed reduced activity and the minus-stranded viral RNA in the replicase became more accessible to ribonuclease when derived from vps23Δ or vps24Δ yeast, suggesting that the protection of the viral RNA is compromised within the replicase complex assembled in the absence of ESCRT proteins. The recruitment of ESCRT proteins is needed for the precise assembly of the replicase complex, which might help the virus evade recognition by the host defense surveillance system and/or prevent viral RNA destruction by the gene silencing machinery. Plus-stranded RNA viruses, which are important pathogens of humans, animals and plants, replicate in infected cells by assembling viral replicase complexes consisting of viral- and host-coded proteins. In this paper, we show that a group of host factors called ESCRT proteins (endosomal sorting complexes required for transport) play important roles in tombusvirus replication. The expression of dominant negative mutants of ESCRT factors inhibited virus replication in the plant host, suggesting that tombusviruses co-opt selected ESCRT proteins for the assembly of the viral replicase complex. In addition, we show direct interaction between the viral p33 replication protein and Vps23p ESCRT-I and Bro1p accessory ESCRT factors. The interaction with p33 leads to the recruitment of Vps23p to the peroxisomes, the sites of tombusvirus replication. We also showed that the viral RNA within the viral replicase complex became more sensitive to ribonuclease in the absence of ESCRT factors, suggesting that the protection of the viral RNA is compromised within the replicase complex assembled in the absence of ESCRT proteins. Intriguingly, the host ESCRT factors also affect the budding of several enveloped viruses, intracellular transport of proteins and cytokinesis. Overall, this work demonstrates that a plus-stranded RNA virus uses the endosomal sorting pathway in a unique way.
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影响因子：
3.7
作者：
Li, Zhenghe;Pogany, Judit;Panavas, Tadas;Xu, Kai;Esposito, Anthony M.;Kinzy, Terri Goss;Nagy, Peter D.
通讯作者：
Nagy, Peter D.
影响因子：
3.7
作者：
Nagy, PD;Pogany, J
通讯作者：
Pogany, J
DOI：
10.1073/pnas.89.21.10208
发表时间：
1992-11-01
影响因子：
11.1
作者：
DOLJA, VV;MCBRIDE, HJ;CARRINGTON, JC
通讯作者：
CARRINGTON, JC
影响因子：
7.2
作者：
Liu, YL;Schiff, M;Dinesh-Kumar, SP
通讯作者：
Dinesh-Kumar, SP
影响因子：
3.7
作者：
Nagy, PD;Pogany, J
通讯作者：
Pogany, J