Li Xiangyang, Zhang Weiying, Ding Yan, Wang Zhenchao, Wu Zengxue, Yu Lu, Hu Deyu, Li Pei, Song Baoan
State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Huaxi District, Guiyang 550025, PR China.
State Key Laboratory Breeding Base of Green Pesticide and Agricultural Bioengineering, Key Laboratory of Green Pesticide and Agricultural Bioengineering, Ministry of Education, Guizhou University, Huaxi District, Guiyang 550025, PR China.
Protein Expr Purif. 2015 Jul;111:98-104. doi: 10.1016/j.pep.2015.04.003. Epub 2015 Apr 11.
Southern rice black-streaked dwarf virus (SRBSDV) P9-1 is involved in viroplasm formations in the SRBSDV-infected plants and insects. During infection, SRBSDV P9-1 is an important protein. However, the function characterization of P9-1 octamers in vitro and in yeast is still obscure. In this study, the secondary and 3D structure of SRBSDV P9-1 was predicted, then SRBSDV P9-1 was expressed and analyzed in vitro, a size exclusion chromatography assay showed that P9-1 had the ability to form octamers in vitro. Mutational analysis of terminal residues showed that the C-terminal arm (residues 324-347) of P9-1 was importance for the formation of octamers. Furthermore, a yeast two-hybrid assay showed that there were strong interactions between the full-length P9-1s, after deleting the C-terminal arm of P9-1, the interactions between the truncated P9-1s were disappeared in yeast. Collectively, the non-structural protein P9-1 played an essential role in self-interact viroplasm formation in vitro and in yeast, and the C-terminal arm region of P9-1 was important for assembling octamers in vitro.
南方水稻黑条矮缩病毒(SRBSDV)P9-1参与了受SRBSDV感染的植物和昆虫中病毒质的形成。在感染过程中,SRBSDV P9-1是一种重要的蛋白质。然而,P9-1八聚体在体外和酵母中的功能特性仍不清楚。在本研究中,预测了SRBSDV P9-1的二级结构和三维结构,然后在体外表达并分析了SRBSDV P9-1,尺寸排阻色谱分析表明P9-1在体外具有形成八聚体的能力。末端残基的突变分析表明,P9-1的C末端臂(残基324-347)对八聚体的形成很重要。此外,酵母双杂交分析表明全长P9-1之间存在强相互作用,删除P9-1的C末端臂后,截短的P9-1之间的相互作用在酵母中消失。总的来说,非结构蛋白P9-1在体外和酵母中自我相互作用的病毒质形成中起重要作用,并且P9-1的C末端臂区域对体外组装八聚体很重要。