Interactions of highly pathogenic avian influenza H5N1 nucleoprotein with NF-κB-inducing kinase(NIK)
10.7644/j.issn.1674-9960.2017.04.002
- VernacularTitle:高致病性禽流感H5N1NP蛋白与NIK蛋白相互作用初步研究
- Author:
Kaiwu LI
;
Ting SONG
;
Wei ZHOU
;
Hongmei DAI
;
Hongguang REN
;
Jing ZHOU
;
Yuan JIN
;
Mingda HU
;
Zhisong HUANG
;
Junjie YUE
;
Long LIANG
- Keywords:
influenza in birds;
influenza A virus;
H5N1 subtype;
nucleoprotein;
NF-κB-inducing kinase;
protein-protein interaction
- From:
Military Medical Sciences
2017;41(4):256-259
- CountryChina
- Language:Chinese
-
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.
