[轮状病毒NSP1蛋白泛素连接酶活性的研究]
[Study on the ubiquitin ligase activity of rotavirus NSP1 protein].
作者信息
Qin Lan, Lei Xiao-Bo, Ren Li-Li, Wang Jian-Wei, Hong Tao
机构信息
State Key Laboratory of Molecular Virology and Genetic Engineering, Institute of Pathogen Biology, Peking Union Medical College & Chinese Academy of Medical Sciences, Beijing 100730.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Dec;24(6):451-4.
OBJECTIVE
To confirm the activity of non structural protein 1 (NSP1) of Rotavirus (RV) as E3 ubiquitin ligase by experiments and to provide some clues for NSP1 on the pathogenic mechanisms and replication of RV.
METHODS
The whole gene and RING deleted mutation gene of NSP1 were coloned into pEGFPC1 expression plasmid, and transfected into human embryonic kidney (HEK) 293 FT cells with pBlue-Script-HA-Ubiquitin. The expression of proteins were proved by using con-focal microscope and western blotting. The ubiquination of proteins were detected by co-immunoprecite.
RESULTS
The cellular proteins of HEK293FT are ubiquinated by NSP1 protein and NSP1 protein was self-ubiquinated also.
CONCLUSIONS
It revealed that RV NSP1 had the activity of E3 ubiquitin ligase and it may play a role on the modulate mechanisms of ubiquination.
目的
通过实验证实轮状病毒（RV）非结构蛋白1（NSP1）作为E3泛素连接酶的活性，为NSP1在RV致病机制和复制方面提供线索。
方法
将NSP1的全长基因及RING结构域缺失的突变基因克隆至pEGFPC1表达质粒中，并与pBlue-Script-HA-泛素一起转染至人胚肾（HEK）293 FT细胞。利用共聚焦显微镜和蛋白质印迹法验证蛋白质的表达。通过免疫共沉淀检测蛋白质的泛素化情况。
结果
HEK293FT细胞的蛋白质被NSP1蛋白泛素化，且NSP1蛋白自身也发生了泛素化。
结论
结果表明RV NSP1具有E3泛素连接酶活性，可能在泛素化调控机制中发挥作用。
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