细胞核受体结合蛋白1通过促进乙肝病毒核心启动子活性增强病毒复制  

Nuclear receptor binding protein 1 enhances the activity of Hepatitis B virus core promoter to promote the replication of Hepatitis B virus

作　　者：周召 鲁凤民[1] Zhou Zhao;Lu Fengmin(Department of Microbiology and Infectious Disease Center,School of Basic Medical Sciences,Peking University Health Science Center,Beijing 100191,China)

机构地区：[1]北京大学基础医学院病原生物学系暨感染病研究中心,100191

出　　处：《国际病毒学杂志》2024年第5期371-375,共5页International Journal of Virology

基　　金：国家自然科学基金项目(82372235)。

摘　　要：目的 探究假激酶细胞核受体蛋白1(nuclear receptor binding protein 1,NRBP1)对乙型肝炎病毒(hepatitis B virus,HBV)复制的作用及相关机制.方法 构建NRBP1外源表达质粒和利用siRNA敲减NRBP1,在瞬转prcccDNA/pCMV-Cre重组质粒系统的慢性乙型肝炎病毒细胞模型中瞬时表达NRBP1或敲减NRBP1.利用实时荧光定量PCR实验和化学发光实验分别检测细胞上清中HBVDNA、HBeAg以及HBsAg的水平变化,反转录-实时荧光定量PCR法检测细胞内HBV RNA含量,western blot实验检测细胞内HBV核心蛋白含量.双荧光素酶报告基因实验检测NRBP1对核心启动子(core promoter,CP)活性的影响.结果 外源表达NRBP1能够使细胞上清中的HBV DNA、HBeAg和HBsAg含量上升.同时外源表达NRBP1也能够使细胞内的core蛋白含量显著上升,使胞内各型HBV RNA含量显著上升.外源表达NRBP1能够显著激活HBV CP启动子活性,且能够显著激活CREB2和c-Jun转录因子的活性.结论 NRBP1促进HBV的转录和复制,可能与其能够上调HBV相关转录因子CREB2和c-Jun的活性有关.Objective To investigate the effect of nuclear receptor binding protein 1(NRBP1)as a pseudokinase on hepatitis B virus(HBV)replication and the related mechanism.Methods The exogenous expression plasmid of NRBP1 was constructed and siRNA was used to knock-down NRBP1.NRBP1 was transient expressed or knock-down in the cell model of chronic heptitis B virus infection with prcccDNA/pCMV-Cre recombinant plasmid transient transfection system.Real-time fluorescence quantitative PCR and chemiluminescence assay were used to detect the changes in the levels of HBV DNA,HBeAg and HBsAg in the supernatant of cell culture.Reverse transcription real-time fluorescence quantitative PCR and western blot were used to detect HBV RNA content and HBV core protein in cells,respectively.Dual luciferase reporter gene assay was used to measure the effect of NRBP1 on core promoter(CP)activity.Results Exogenous expression of NRBP1 can increase the content of HBV DNA,HBeAg and HBsAg in the supernatant of cell culture.It also increased significantly the levels of core protein,within cells,resulting in elevation of various types of HBV RNA.Exogenous expression of NRBP1 can significantly activate HBV core promoter as well as transcription factors CREB2 and c-jun.Conclusions NRBP1 can promote HBV transcription and replication,which may be related to the ability of up-regulating HBV-related transcription factors,e.g.CREB2 and c-jun.

关 键 词：乙型肝炎病毒 细胞核受体蛋白1 病毒复制 病毒转录 

分 类 号：R512.62[医药卫生—内科学]