宿主蛋白TRIB2与流感病毒NP蛋白相互作用的研究  

Study on the interaction between influenza virus NP protein and host protein TRIB2

作　　者：周圆[1] 赵玉辉[1] 张杰[1] 赵青青[1] 周陈陈 王广文[1] 梁立滨[1] 李俊平[1] 姜丽[1] 陈化兰[1] 李呈军[1] ZHOU Yuan ZHAO Yu-hui ZHANG Jie ZHAO Qing-qing ZHOU Chen-chen WANG Guang-wen LIANG Li-bin LI Jun-ping JIANG Li CHEN Hua-lan LI Cheng-jun＊(Animal Influenza Key Laboratory of the Ministry of Agriculture, State Key Laboratory of Veterinary Bioteehnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Science, Harbin 150069, China)

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/农业部动物流感重点开放实验室,黑龙江哈尔滨150069

出　　处：《中国预防兽医学报》2017年第2期85-89,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31472215;31521005)

摘　　要：流感病毒的核衣壳蛋白(NP)是病毒粒子中重要的结构蛋白,参与流感病毒生命周期的多个过程。为筛选与流感病毒NP相互作用的宿主蛋白,本研究应用酵母双杂交技术(Y2H)技术从A549细胞cDNA文库中筛选到与其相互作用的宿主蛋白Tribble 2(TRIB2),并通过免疫共沉淀技术(Co-IP)试验和GST pull down试验进一步证实NP与TRIB2之间存在特异性的直接相互作用。此外,利用激光共聚焦试验证实NP与TRIB2共定位于A549细胞的细胞质内。在A549细胞中过表达TRIB2蛋白后,能够降低流感病毒的滴度,表明TRIB2具有抑制流感病毒的复制功能。本实验为研究流感病毒的复制机理奠定了基础,完善了病毒与宿主相互作用网络。Nucleoprotein （NP） is one of the most abundant structural proteins of influenza virus particle, and is involved in multiple processes of the virus life cycle. To screen for host interacting partners of NP protein, we applied a yeast two-hybrid strategy by using NP as bait. TRIB2 was identified and its interaction with NP was further demonstrated by co-immunopr- ecipitation （Co-IP） and GST pull-down experiments. Moreover, confocal microscopy showed that NP and TRIB2 were colocalized in the cytoplasm of virus-infected A549 cells. We further found that overexpression of TRIB2 inhibited the virus titer. Our data showed that TRIB2 protein play a role in the replication of influenza virus, laying a foundation for further studies of the regulation mechanisms of the virus life cycle.

关 键 词：酵母双杂交 NP蛋白 TRIB2蛋白 流感病毒 

分 类 号：S852.65[农业科学—基础兽医学]