Polydnaviruses associated with ichneumonid parasitoid wasps (Ichnoviruses) encode large numbers of genes, often in multigene families. The Ichnovirus Vinnexin gene family, which is expressed in parasitized lepidopteran larvae, encodes homologues of Innexins, the structural components of insect gap junctions. Here, we have examined intracellular behaviours of the Campoletis sonorensis Ichnovirus (CsIV) Vinnexins, alone and in combination with a host Innexin orthologue, Innexin2 (Inx2). QRT‐PCR verified that transcription of CsIV vinnexins occurs contemporaneously with inx2, implying co‐occurrence of Vinnexin and Inx2 proteins. Confocal microscopy demonstrated that epitope‐tagged VinnexinG (VnxG) and VinnexinQ2 (VnxQ2) exhibit similar subcellular localization as Spodoptera frugiperda Inx2 (Sf‐Inx2). Surface biotinylation assays verified that all three proteins localize to the cell surface, and cytochalasin B and nocodazole that they rely on actin and microtubule cytoskeletal networks for localization. Immunomicroscopy following co‐transfection of constructs indicates extensive co‐localization of Vinnexins with each other and Sf‐Inx2, and live‐cell imaging of mCherry‐labelled Inx2 supports that Vinnexins may affect Sf‐Inx2 distribution in a Vinnexin‐specific fashion. Our findings support that the Vinnexins may disrupt host cell physiology in a protein‐specific manner through altering gap junctional intercellular channel communication, as well as indirectly by affecting multicellular junction characteristics.

中文翻译：

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Polydnavirus内毒素同系物的细胞内动力学。

与棘孢类寄生性类黄蜂（鱼类病毒）有关的多角膜病毒编码大量基因，通常是多基因家族。在寄生的鳞翅目幼虫中表达的支气管病毒Vinnexin基因家族编码昆虫间隙连接的结构成分Innexins的同系物。在这里，我们已经检查了单独或与宿主Innexin直向同源物Innexin2（Inx2）结合使用的Campoletis sonorensis线虫病毒（CsIV）Vinnexins的细胞内行为。QRT‐PCR证实CsIV vinnexins的转录与inx2同时发生，表示Vinnexin和Inx2蛋白同时存在。共聚焦显微镜显示，表位标记的VinnexinG（VnxG）和VinnexinQ2（VnxQ2）表现出与节食夜蛾Inx2 （Sf- Inx2）类似的亚细胞定位。表面生物素化测定法证实，所有三种蛋白质均位于细胞表面，而细胞松弛素B和诺考达唑则依赖肌动蛋白和微管细胞骨架网络进行定位。共转染构建物后的免疫镜检查显示Vinnexins与Sf -Inx2相互之间广泛共定位，mCherry标记的Inx2的活细胞成像支持Vinnexins可能影响Sf‐以Vinnexin特定的方式分发Inx2。我们的发现支持Vinnexins可以通过改变缝隙连接细胞间通道之间的通讯，以及通过影响多细胞连接特性而间接地以蛋白质特异性的方式破坏宿主细胞的生理。