风疹病毒衣壳蛋白与宿主细胞相互作用蛋白的初步筛选及分析  

Screening and Analyses of Proteins Interacting with the Capsid Protein of the Rubella Virus in Host Cells

作　　者：韩月雯 吴瑞[1] 马超锋[1] 李园园 HAN Yuewen;WU Rui;MA Chaofeng;LI Yuanyuan(Viral Diseases Laboratory of Xi’an Center for Disease Control and Prevention,Xi’an 710054,China;Dalian Rehabilitation Center of Joint Logistic Force of PLA,Dalian 116013,China)

机构地区：[1]西安市疾病预防控制中心病毒病检验科,西安710054 [2]联勤保障部队大连康复疗养中心,大连116013

出　　处：《病毒学报》2021年第5期1074-1078,共5页Chinese Journal of Virology

基　　金：陕西省自然科学基础研究计划面上项目(项目号:2017JM8101),题目:风疹病毒蛋白与宿主蛋白相互作用关系及功能学研究。

摘　　要：风疹病毒(Rubella virus,RV)的衣壳蛋白(Capsid protein,CP)不仅是病毒颗粒的重要组成部分,而且还可以通过与宿主蛋白之间的相互作用来调控病毒的转录和复制。为了系统研究衣壳蛋白与宿主蛋白之间的相互作用关系,我们从RV基因组中克隆获得衣壳蛋白基因序列,将该序列导入含有eXact⁃6His串联亲和纯化标签的慢病毒表达载体中,并构建了稳定表达eXact⁃6His⁃Capsid融合蛋白的293T细胞系。通过eXact和6His标签的两次亲和纯化获得衣壳蛋白相互作用蛋白复合物,质谱检测并筛选后发现22个可能与衣壳蛋白相互作用的宿主蛋白。随后构建衣壳蛋白的相互作用网络并进行功能学分析,发现其相互作用蛋白主要参与病毒感染、RNA剪切、细胞凋亡及酶相关通路等过程。The capsid protein(Capsid protein,CP)of the rubella virus(Rubella virus,RV)is not only an important component of the virus particle,but can also regulate the transcription and replication of the RV through interactions with host proteins.We wished to systematically study the interactions between the capsid protein of the RV and host proteins.First,the sequence of the capsid gene was cloned from the RV genome.Then,the sequence was imported into the lentivirus expression vector containing the eXact⁃6His tandem affinity purification tag.A 293T cell line stably expressing the eXact⁃6His⁃Capsid recombinant protein was constructed.Then,complexes of interacting proteins were purified by the eXact⁃6His tandem affinity tag.After detection and screening using mass spectrometry,22 proteins that interacted with the capsid protein of the RV were obtained,and a protein–protein interaction network mapped.Bioinformatics analysis found that these interacting proteins were involved in viral infection,RNA splicing,apoptosis,and enzyme⁃related pathways.This work provides a basis for a detailed map of interactions between the capsid protein of the RV and host proteins,and provides clues for the in⁃depth study of RV infection/pathogenesis.

关 键 词：风疹病毒(RV) 衣壳蛋白(CP) 相互作用蛋白 串联亲和纯化 质谱 功能分析 

分 类 号：R34[医药卫生—基础医学]