Incorporation of the Kaposi's sarcoma-associated herpesvirus capsid vertex-specific component (CVSC) into self-assembled capsids.
Incorporation of the Kaposi's sarcoma-associated herpesvirus capsid vertex-specific component (CVSC) into self-assembled capsids.
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DOI：
10.1016/j.virusres.2017.04.016
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发表时间：
2017-05-15
期刊：
影响因子：
5
通讯作者：
Desai PJ
中科院分区：
文献类型：
--
作者：
Grzesik P;MacMath D;Henson B;Prasad S;Joshi P;Desai PJ
疱疹病毒衣壳的自组装可以在外源表达系统中完成，前提是所有六种衣壳蛋白都存在。我们利用杆状病毒表达系统在昆虫细胞中展示了二十面体卡波西肉瘤相关疱疹病毒（KSHV）衣壳的组装。利用这个自组装系统，我们研究了是否可以添加额外的衣壳相关蛋白，并确定它们在组装衣壳中的结合情况。我们选择由开放阅读框（orf） 19和32编码的衣壳顶点特异性成分（CVSC）蛋白来测试这一点。这种复合物位于衣壳顶点，对疱疹病毒感染细胞的衣壳成熟很重要。共免疫沉淀试验初步证实ORF19和ORF32之间存在双分子相互作用。这两种蛋白还沉淀了衣壳壳的三联体蛋白（ORF26和ORF62）以及主要的衣壳蛋白（ORF25）。衣壳免疫沉淀实验显示ORF19和ORF32结合到组装的衣壳中。类似的实验也表明，每种蛋白质的结合都是相互独立的。这些研究揭示了KSHV CVSC与衣壳壳的生物化学相互作用。
Self-assembly of herpesvirus capsids can be accomplished in heterologous expression systems provided all six capsid proteins are present. We have demonstrated the assembly of icosahedral Kaposi’s sarcoma-associated herpesvirus (KSHV) capsids in insect cells using the baculovirus expression system. Using this self-assembly system we investigated whether we could add additional capsid associated proteins and determine their incorporation into the assembled capsid. We chose the capsid vertex-specific component (CVSC) proteins encoded by open reading frames (ORFs) 19 and 32 to test this. This complex sits on the capsid vertex and is important for capsid maturation in herpesvirus-infected cells. Co-immunoprecipitation assays were used to initially confirm a bi-molecular interaction between ORF19 and ORF32. Both proteins also precipitated the triplex proteins of the capsid shell (ORF26 and ORF62) as well as the major capsid protein (ORF25). Capsid immunoprecipitation assays revealed the incorporation of ORF19 as well as ORF32 into assembled capsids. Similar experiments also showed that the incorporation of each protein occurred independent of the other. These studies reveal biochemically how the KSHV CVSC interacts with the capsid shell.
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