Characterization and interactome study of white spot syndrome virus envelope protein VP11.
Characterization and interactome study of white spot syndrome virus envelope protein VP11.
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DOI：
10.1371/journal.pone.0085779
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Chang YS
中科院分区：
文献类型：
--
作者：
Liu WJ;Shiung HJ;Lo CF;Leu JH;Lai YJ;Lee TL;Huang WT;Kou GH;Chang YS
白色斑点综合征病毒（WSSV）是一种大型包膜病毒。WSSV病毒颗粒由围绕其核心DNA的三个结构层组成：外被膜、被膜和核衣壳。在这里，我们的特点是WSSV结构蛋白VP 11（WSSV 394，GenBank登录号AF 440570），并使用相互作用组的方法来分析这种蛋白质和其他WSSV和宿主蛋白的阵列之间的可能关联。时间转录分析表明vp 11是一个早期基因。完整病毒颗粒的Western印迹杂交和病毒成分的分级分离以及免疫电子显微镜显示VP 11是一种包膜蛋白。膜拓扑学软件预测VP 11是一种跨膜蛋白，其N端具有高度疏水的跨膜结构域。基于对VP 11转染的Sf 9细胞进行的免疫荧光测定和对病毒体的胰蛋白酶消化分析，我们得出结论，与拓扑软件预测相反，该蛋白质的C-末端实际上在病毒体内部。酵母双杂交筛选结合免疫共沉淀试验发现，VP 11直接与至少12个其他WSSV结构蛋白以及本身相互作用。寡聚化试验进一步表明VP 11可形成二聚体。VP 11也是第一个报道的WSSV结构蛋白与主要核衣壳蛋白VP 664相互作用。
White spot syndrome virus (WSSV) is a large enveloped virus. The WSSV viral particle consists of three structural layers that surround its core DNA: an outer envelope, a tegument and a nucleocapsid. Here we characterize the WSSV structural protein VP11 (WSSV394, GenBank accession number AF440570), and use an interactome approach to analyze the possible associations between this protein and an array of other WSSV and host proteins. Temporal transcription analysis showed that vp11 is an early gene. Western blot hybridization of the intact viral particles and fractionation of the viral components, and immunoelectron microscopy showed that VP11 is an envelope protein. Membrane topology software predicted VP11 to be a type of transmembrane protein with a highly hydrophobic transmembrane domain at its N-terminal. Based on an immunofluorescence assay performed on VP11-transfected Sf9 cells and a trypsin digestion analysis of the virion, we conclude that, contrary to topology software prediction, the C-terminal of this protein is in fact inside the virion. Yeast two-hybrid screening combined with co-immunoprecipitation assays found that VP11 directly interacted with at least 12 other WSSV structural proteins as well as itself. An oligomerization assay further showed that VP11 could form dimers. VP11 is also the first reported WSSV structural protein to interact with the major nucleocapsid protein VP664.
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