杆状病毒经口感染因子之间相互作用分析  

Analysis of the Interactions Among Baculovirus Per Os Infectivity Factors (PIFs)

作　　者：郑秦[1] 范颖 王海萍[1] 许伟凡 孔祥硕 吴小锋[1] 

机构地区：[1]浙江大学动物科学学院

出　　处：《病毒学报》2018年第1期85-93,共9页Chinese Journal of Virology

基　　金：国家自然科学基金项目(项目号:31472146),题目:家蚕BmNPV出芽型病毒囊膜蛋白GP64受体蛋白的分子鉴定;国家自然科学基金项目(项目号:31772675),题目:家蚕BmNPV沿气管体内传播的分子机制~~

摘　　要：昆虫杆状病毒包涵体衍生型病毒（Occlusion-derived virus,ODV）被幼虫食下后通过感染中肠上皮细胞引发全身感染,这个过程被称为经口感染。多种ODV囊膜蛋白在经口感染中发挥关键作用,它们被称为经口感染因子（PIFs）。研究表明7种PIFs：P74、PIF1、PIF2、PIF3、PIF4、PIF6、P95（Ac83）和2种ODV特异性蛋白,Ac5和Ac108,与ODV表面复合物相关,该复合物被称为PIF复合物。阐明杆状病毒PIFs之间,尤其是复合物各个成分之间的相互作用特点是了解经口感染因子功能的关键。本研究以杆状病毒模式种-苜蓿银纹夜蛾核型多角体病毒（Autographa californica multiple nucleopolyhedrovirus,AcMNPV）为研究对象,利用酵母双杂交和双分子荧光互补技术探究了与PIF复合物相关的9种蛋白之间相互作用的特点。我们鉴定到了6对相互作用,分别为PIF1-PIF2、PIF1-PIF3、PIF1-PIF4、PIF1-P95、PIF2-PIF3和PIF3-PIF4,但是没有鉴定到P74、PIF6、Ac5和Ac108参与相互作用。其中PIF1-PIF3、PIF2-PIF3和PIF1-P95为强相互作用,其他相互作用为中等强度相互作用。根据这些结果我们推测PIF1是PIF复合物的中心,其他成分都和PIF1存在相互作用。最后,我们对目前为止已报道的PIF之间相互作用进行了总结,包括10对不同蛋白之间的相互作用：PIF1-PIF2、PIF1-PIF3、PIF1-PIF4、PIF1-P95、PIF2-PIF3、PIF3-PIF4、ODV-E56-PIF1、ODV-E56-PIF2、ODV-E56-PIF3和ODV-E56-P74,以及2对自相互作用：PIF3-PIF3和ODV-E56-ODV-E56。总之,本研究探索了PIF复合物中蛋白质的相互作用特点,揭示了复合物中各成分如何通过相互作用有机地结合在一起,推进了对该复合物的认识。The occlusion-derived viruses（ODVs）of baculoviruses establish the first round of infection within the midgut cells of the larval host after ingestion of occlusion bodies.Several ODV envelope proteins have been shown to be essential for oral infection：these are called per os infectivity factors（PIFs）.Nine PIFs have been identified：P74,PIFs 1~6,Ac83 and Ac110.At least seven PIFs（P74,PIFs 1~4,P95（Ac83）and PIF6）together with two other ODV-specific proteins（Ac5 and Ac108）have been reported to form complexes on ODV surface.Here,to understand the interactions of these PIFs,the direct protein–protein interactions of the nine components of the Autographa californica multiple nucleopolyhedrovirus PIF complex were investigated using yeast two-hybrid screening combined with the bimolecular fluorescence complementation assay.Six direct interactions were identified：PIF1-PIF2,PIF1-PIF3,PIF1-PIF4,PIF1-P95,PIF2-PIF3 and PIF3-PIF4.No direct interaction was identified for P74,PIF6,Ac5 and Ac108.Among these interactions,PIF1-PIF3,PIF2-PIF3 and PIF1-P95 were strong,whereas the others appeared to be less robust.According to these results,we speculated that PIF1 is the core of the PIF complex that other components can interact with it.Furthermore,a summary of the protein-protein interactions of PIFs reported so far,which comprise ten reciprocal interactions（PIF1-PIF2,PIF1-PIF3,PIF1-PIF4,PIF1-P95,PIF2-PIF3,PIF3-PIF4,ODV-E56-PIF1,ODV-E56-PIF2,ODV-E56-PIF3 and ODV-E56-P74）and two self-interactions（PIF3-PIF3 and ODV-E56-ODV-E56）is presented.In conclusion,we explored the characteristics of PIF complexes and elucidated how the proteins in PIF complexes interact with each other,which will help us to better understand this complex of baculovirus and it role in viral infection.

关 键 词：杆状病毒 包涵体衍生型病毒(ODV) 经口感染因子(PIFs) 相互作用 酵母双杂交 双分子荧光互补 

分 类 号：S476.13[农业科学—农业昆虫与害虫防治]