军事医学2017，Vol.41Issue(4)：256-259,4.DOI:10.7644/j.issn.1674-9960.2017.04.002
高致病性禽流感H5N1NP蛋白与NIK蛋白相互作用初步研究
Interactions of highly pathogenic avian influenza H5N1 nucleoprotein with NF-κB-inducing kinase(NIK)
摘要
Abstract
Objective To study the interactions between the highly pathogenic avian influenza H5N1 nucleoprotein (H5N1 NP) and NF-κB-inducing kinase (NIK),and to reveal the effect of H5N1 NP on NIK-induced NF-κB transcriptional activity.Methods The gene encoding NIK protein was amplified by RT-PCR from total RNA of HeLa cell line.Eukaryotic expression plasmid pCMV-Myc-NIK and prokaryotic expression plasmid pGEX-4T-1-NP (GST-NP) were constructed by cloning from HeLa cell cDNA and pcDNA3-Flag-NP vector,respectively.Co-immunoprecipitation (co-IP) and GST pull-down were used to test the interactions between H5N1 NP and NIK.Dual-luciferase reporter gene analysis system was used to test the effect of H5N1 NP on NIK-induced NF-κB transcriptional activity.Results Co-IP and GST pull-down showed that pCMV-Myc-NIK and pGEX-4T-1-NP (GST-NP) could express Myc tagged NIK protein and GST tagged NP protein in HEK293T cells and E.coli,respectively,and that H5N1 NP was associated with NIK in vivo and in vitro.Dual-luciferase reporter gene analysis suggested that H5N1 could inhibit NIK-induced NF-κB transcriptional activity.Conclusion H5N1 NP interacts with NIK and inhibits NIK-induced NF-κB transcriptional activity.This finding can facilitate further study of H5N1.关键词
禽流感/流感病毒A型,H5N1亚型/核蛋白/NF-κB诱导激酶/蛋白质相互作用Key words
influenza in birds/influenza A virus/H5N1 subtype/nucleoprotein/NF-κB-inducing kinase/protein-protein interaction分类
医药卫生引用本文复制引用
李凯武,岳俊杰,梁龙,宋婷,周围,戴红梅,任洪广,周静,靳远,胡明达,黄志松..高致病性禽流感H5N1NP蛋白与NIK蛋白相互作用初步研究[J].军事医学,2017,41(4):256-259,4.基金项目
国家自然科学基金资助项目(31400046) （31400046）
