宿主蛋白LSm1-7复合体在登革病毒RNA复制中的作用研究  

The role of host factor LSm1-7 complex in dengue virus RNA replication

作　　者：董阳超[1] 雷迎峰[1] 丁天兵[1] 时莹[1] 潘鹭翔[1] 王晓霞[2] 张泽信[2] 徐志凯[1] 

机构地区：[1]第四军医大学微生物学教研室,陕西西安710032 [2]第四军医大学西京医院麻醉科,陕西西安710032

出　　处：《热带医学杂志》2013年第1期13-16,F0004,共5页Journal of Tropical Medicine

摘　　要：目的探讨LSm1-7复合体在登革病毒(DENV)RNA复制中的作用。方法 DENV感染HepG2细胞后,在不同时间点(2、24、36、48h)收集细胞和提取总mRNA,实时定量PCR(RT-qPCR)分析LSm1表达水平;将LSm1的siRNA和非特异对照siRNA两次转染HepG2细胞后感染DENV-2,24h后收集细胞,提取总mRNA,RT-qPCR分析LSm1表达水平与病毒RNA表达水平;利用LSm1抗体与dsRNA抗体对DENV感染细胞进行免疫荧光染色,激光共聚焦分析LSm1与dsRNA共定位情况。结果与2h比较,随着时间的推移,DENV-2 RNA和LSm1 RNA水平逐渐升高;与对照组siNC比较,siLSm1组LSm1 RNA水平明显降低,沉默效率达78.2%;与对照组siNC比较,siLSm1组DENV RNA含量降低35.6%;LSm1-7复合体在DENV感染HepG2细胞后募集到核周围,与DENV dsRNA共定位。结论 LSm1-7复合体可能作为DENV复制复合物的组成部分,正调控DENV RNA的复制。Objective To investigate the roles of host protein LSml-7 complex in DENV RNA replication. Methods mRNA was isolated from DENV infected HepG2 cells at different time point（2,24,36,48 h） of post-infection, and the level of DENV RNA and LSml RNA was analysed by RT-qPCR. HepG2 cells were transfected with specific siRNA for LSml and nontargeting siRNA and siNC respectively,followed by infection with DENV.The mRNA was isolated at 24 h p. i. and analyzed by RT-qPCR to determine the RNA level of DENV and LSml. Laser scanning eonfoeal microscopy was performed to observe the localization of LSml protein and DENV dsRNA in DENV infected HepG2 cells. Results The levels of DENV RNA and LSml RNA were increased compared to 2 h p.i. during the course of DENV infection, siRNA- mediated silencing resulted in a specific 78.2% reduction of LSml RNA when using the nontargeting siNC as a negative control, and down-regulation of LSml resulted in a marked reduction of the DENY RNA level by 35.6%. LSml-7 complex distributed and colocalized with DENV dsRNA Conclusion LSml-7 complex may act as the component regulators for DENV RNA replication. around nucleolous in the DENV infected HepG2 cells. of DENV replication machinery and is one of the positive

关 键 词：登革病毒 宿主蛋白 LSm1-7复合体 RNA复制 

分 类 号：R373.3[医药卫生—病原生物学]