Identification of insect genes involved in baculovirus AcMNPV entry into insect cells
Identification of insect genes involved in baculovirus AcMNPV entry into insect cells
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DOI：
10.1016/j.virol.2018.10.022
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发表时间：
2019-01-15
期刊：
影响因子：
3.7
通讯作者：
Blissard, Gary W.
中科院分区：
文献类型：
--
作者：
Hodgson, Jeffrey J.;Buchon, Nicolas;Blissard, Gary W.
苜蓿银纹夜蛾核型多角体病毒（Autographa californica multiple nucleopolyhedrovirus，AcMNPV）是一种能在鳞翅目昆虫细胞中复制的模式DNA病毒，能有效地进入多种非宿主细胞。AcMNPV的出芽病毒粒子通过内吞作用进入细胞并运输到细胞核，在细胞核中病毒启动基因表达和基因组复制。虽然贩运核衣壳肌动蛋白推进已被详细研究，其他重要组成部分的贩运过程中进入仍然知之甚少。我们使用了一个重组AcMNPV病毒表达的EGFP报告基因结合RNAi屏幕在果蝇DL1细胞，以确定参与AcMNPV进入宿主蛋白。RNAi筛选靶向参与囊泡运输的86个基因，包括编码VPS和ESCRT蛋白、Rab GTP酶、外囊蛋白和网格蛋白衔接蛋白的基因。我们确定了24个基因所需的有效病毒进入和报告基因表达，和4个基因，似乎限制病毒进入。
The baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is a model enveloped DNA virus that infects and replicates in lepidopteran insect cells, and can efficiently enter a wide variety of non-host cells. Budded virions of AcMNPV enter cells by endocytosis and traffic to the nucleus where the virus initiates gene expression and genome replication. While trafficking of nucleocapsids by actin propulsion has been studied in detail, other important components of trafficking during entry remain poorly understood. We used a recombinant AcMNPV virus expressing an EGFP reporter in combination with an RNAi screen in Drosophila DL1 cells, to identify host proteins involved in AcMNPV entry. The RNAi screen targeted 86 genes involved in vesicular trafficking, including genes coding for VPS and ESCRT proteins, Rab GTPases, Exocyst proteins, and Clathrin adaptor proteins. We identified 24 genes required for efficient virus entry and reporter expression, and 4 genes that appear to restrict virus entry.