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Identification of a novel host protein interacting with the structural protein VP2 of deformed wing virus by yeast two-hybrid screening.
Virus Research ( IF 5 ) Pub Date : 2020-07-10 , DOI: 10.1016/j.virusres.2020.198072
Sichao Huang 1 , Dongliang Fei 1 , Yueyu Ma 1 , Chen Wang 1 , Donghui Shi 1 , Kunyang Liu 2 , Ming Li 1 , Mingxiao Ma 1
Affiliation  

Deformed wing virus (DWV) interacting with Varroa destructor is a possible cause of honeybee colony mortality. VP2 is the structural protein of DWV but its function remains unknown. To clarify the function of VP2 and screen for novel binding proteins that interact with VP2, we carried out a membrane protein yeast two-hybrid screening using VP2 as bait. Subsequently, the interaction between VP2 and the host interacting protein [heat shock protein 10 (Hsp10)] was further verified using glutathione S-transferase pull-down assay in vitro and co-immunoprecipitation assay in cells. Furthermore, fluorescence confocal microscopy revealed that VP2 and Hsp10 were mainly co-localized in the cytoplasm. Using real-time polymerase chain reaction, we found that Hsp10 expression in DWV-infected worker honey bees were downregulated compared with that in healthy honey bees. Additionally, we showed that overexpression of VP2 protein could reduce the expression of Hsp10. These results suggest that Hsp10 plays a vital role in host immunity and antiviral effects.



中文翻译:

通过酵母双杂交筛选鉴定与变形翼病毒的结构蛋白VP2相互作用的新型宿主蛋白。

变形翼病毒(DWV)与Varroa破坏因子相互作用是蜜蜂菌落死亡的可能原因。VP2是DWV的结构蛋白,但其功能尚不清楚。为了阐明VP2的功能并筛选与VP2相互作用的新型结合蛋白,我们以VP2为诱饵进行了膜蛋白酵母双杂交筛选。随后,体外使用谷胱甘肽S-转移酶下拉试验和细胞中的免疫共沉淀试验进一步验证了VP2与宿主相互作用蛋白[热激蛋白10(Hsp10)]之间的相互作用。此外,荧光共聚焦显微镜显示VP2和Hsp10主要共定位在细胞质中。使用实时聚合酶链反应,我们发现与健康蜜蜂相比,DWV感染工蜂中的Hsp10表达下调。此外,我们表明VP2蛋白的过表达可以减少Hsp10的表达。这些结果表明,Hsp10在宿主免疫和抗病毒作用中起着至关重要的作用。

更新日期:2020-07-17
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