Liu Weisha, Wang Hao, Yu Fei, Lu Liqun
Key Laboratory of Freshwater Fishery Germplasm Resources, National Pathogen Collection Center for Aquatic Animals, Shanghai Ocean University, Shanghai, 201306, China.
Arch Virol. 2017 Aug;162(8):2375-2380. doi: 10.1007/s00705-017-3354-9. Epub 2017 Apr 18.
Grass carp reovirus (GCRV) causes viral hemorrhagic disease in cultured grass carp. However, there is no effective means of controlling GCRV. Previous studies have shown, by cryoelectron microscopy, that the outer capsid of GCRV is composed of 200 trimers of VP5-VP7 heterodimers. However, confirmation of this interaction between VP5 and VP7 through molecular biochemistry is still lacking. This study characterized the interactions between VP5 and VP7 in vitro. VP5 was shown to interact with VP7 in a commercial yeast-two-hybrid screen. A dot-blot overlay assay was used to show that VP7 binding to VP5 was dose-dependent. Finally, a yeast-two-hybrid approach confirmed interactions between full-length and truncated forms of VP5 and VP7. In conclusion, our results indicate that VP5 and VP7 interact directly in vitro.
草鱼呼肠孤病毒(GCRV)可导致养殖草鱼患病毒性出血病。然而,目前尚无有效的控制GCRV的方法。先前的研究通过冷冻电子显微镜显示,GCRV的外衣壳由200个VP5-VP7异二聚体三聚体组成。然而,通过分子生物化学对VP5和VP7之间这种相互作用的证实仍然缺乏。本研究对VP5和VP7在体外的相互作用进行了表征。在商业酵母双杂交筛选中显示VP5与VP7相互作用。斑点印迹覆盖分析表明VP7与VP5的结合是剂量依赖性的。最后,酵母双杂交方法证实了VP5和VP7全长及截短形式之间的相互作用。总之,我们的结果表明VP5和VP7在体外直接相互作用。