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当前位置: 首页 > 文章 > 猪脑心肌炎病毒3AB蛋白与宿主蛋白RPL11相互作用的鉴定 中国预防兽医学报 2018 (5) 375-380
Position: Home > Articles > Identification of the interaction between encephalomyocarditis virus 3AB protein and host protein 60S ribosomal protein L11 Chinese Journal of Preventive Veterinary Medicine 2018 (5) 375-380

猪脑心肌炎病毒3AB蛋白与宿主蛋白RPL11相互作用的鉴定

作  者：
薛娟;张昆丽;张元峰;黄丽;杨玉莹;翁长江
单  位：
中国农业科学院哈尔滨兽医研究所基础免疫创新团队/兽医生物技术国家重点实验室;长江大学动物科学学院
关键词：
猪脑心肌炎病毒;3AB蛋白;60S核糖体蛋白L11;病毒-宿主相互作用
摘  要：
为了研究与猪脑心肌炎病毒(EMCV)3AB相互作用的宿主蛋白,本实验采用酵母双杂交方法筛选BHK-21细胞cDNA酵母表达文库,获得6个与EMCV 3AB相互作用的宿主蛋白(RFC5、LRWD1、PIH1D1、RPL11、RPS20和LGALS1),经分析显示这些蛋白分别参与DNA复制、有丝分裂和细胞凋亡等过程;酵母共转化试验与免疫共沉淀试验证实EMCV 3AB和RPL11两者之间存在特异性结合;激光共聚焦试验显示EMCV 3AB与RPL11共同过度表达时,二者共定位在细胞核中,而在EMCV感染状态下,3AB与RPL11主要共定位在细胞质中。另外,EMCV感染HeLa细胞能降低细胞中p53的蛋白水平,但3AB并不直接促进p53蛋白降解,表明EMCV 3AB对RPL11-MDM2-p53通路无影响。本研究发现并证实了RPL11与EMCV 3AB之间的相互作用,为进一步研究RPL11蛋白在EMCV复制中的作用奠定了基础。
译  名：
Identification of the interaction between encephalomyocarditis virus 3AB protein and host protein 60S ribosomal protein L11
作  者：
XUE Juan;ZHANG Kun-li;ZHANG Yuan-feng;HUANG Li;YANG Yu-ying;WENG Chang-jiang;College of Animal Science, Yangtze University;Harbin Veterinary Research Institute,Chinese Academy of Agricultural Sciences/State Key Laboratoryof Veterinary Biotechnology;
关键词：
encephalomyocarditis virus;;3AB protein;;RPL11;;virus-host interaction
摘  要：
To identify encephalomyocarditis virus(EMCV) 3AB protein binding partner, 3AB protein was used as "bait" to screen the Baby Hamster Kidney 21(BHK-21) cDNA expression library in Yeast by yeast two-hybrid(Y2H). Six host proteins,RFC5, LRWD1, PIH1D1, RPL11, RPS20 and LGALS1, were identified as 3AB binding partners. In this study, the interaction of RPL11 and 3AB was further verified by Y2 H retest and co-immunoprecipitation(Co-IP) assay. In addition, the localization of RPL11 and 3AB protein was tested by laser confocal assay. We found that HA tagged EMCV 3AB and Flag-RPL11 colocalized in the nucleolar in HeLa cells. However, the colocation of ectopic expression RPL11 and EMCV-3AB changed from nuclear to cytoplasm when EMCV infected He La cells. Moreover, 3AB was unable to directly promote p53 degradation although EMCV infection led to p53 degradation in He La cells. Our findings demonstrated that RPL11 interacted with EMCV 3AB protein and RPL11 changed EMCV 3AB subcellular localization, which will facilitate further study of the role of RPL11 in EMCV replication.

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