Interaction between influenza A virus nucleoprotein and TRIM25 protein
10.3969/j.issn.1002-2694.2025.00.032
- VernacularTitle:A型流感病毒NP蛋白与TRIM25蛋白互作的研究
- Author:
Xin-yan HU
1
;
Qian-yun LIU
;
Le-le AN
;
Qiu-ju LAN
;
Xiao-xia MA
Author Information
1. 西北民族大学生物医学研究中心,生物工程与技术国家民委重点实验室,兰州 730030;西北民族大学生物医学研究中心,甘肃省动物细胞技术创新中心,兰州 730030;西北民族大学,生命科学与工程学院,兰州 730010
- Publication Type:Journal Article
- Keywords:
influenza A virus;
nucleoprotein;
TRIM25;
protein interaction
- From:
Chinese Journal of Zoonoses
2025;41(3):219-226
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed at exploring the interaction between the nucleoprotein(NP)of influenza A virus(IAV)and TRIM25.The physicochemical properties and protein structure of IAV NP protein were analyzed through bioinformatics methods.The interaction between IAV NP and TRIM25 proteins was simulated with molecular docking techniques,and the in-teraction sites were predicted.With the cDNA of the A/Puerto Rico/8/1934(H1N1)PR8 strain as the template,the NP pro-tein was cloned into the eukaryotic expression vector pCMV-C-Flag through PCR amplification,the eukaryotic expression re-combinant plasmid pCMV-Flag-NP was constructed,and the expression was further verified.The protein expression levels of pCMV-Flag-NP and pCMV-HA-TRIM25 were detected at various time periods.The interaction between NP protein and TRIM25 protein was verified by co-immunoprecipitation.The co-localization of NP protein and TRIM25 protein in cells was ob-served with laser confocal microscopy.Bioinformatics analysis revealed that the NP protein consists of 498 amino acids and 20 amino acids,and is an unstable hydrophilic protein.The NP protein has multiple phosphorylation sites,as well as N-glycosyla-tion and O-glycosylation sites,but no transmembrane domain or signal peptide domain.Additionally,the NP protein's second-ary structure consists of a high proportion of alpha-helices and random coils.The molecular docking prediction results indicated that IAV NP interacts with TRIM25 protein and has multiple potential interaction sites,including the 233rd alanine,234th ala-nine,236th lysine,and 440th alanine of the NP protein.After successfully constructing and expressing the IAV NP protein,we verified the interaction between IAV NP and TRIM25 protein by immunoprecipitation and laser confocal microscopy obser-vations.Our results together suggested that the structure of the IAV NP protein is closely related to its function,and its im-portance to the virus is clear.In addition,the interaction between IAV NP and TRIM25 protein may be associated with TRIM25's anti-influenza virus mechanism.Further in-depth research may provide new ideas for anti-influenza virus strategies.
