ADAR2通过编辑MAVS基因的3'UTR降低其在细胞中的表达  

ADAR2 inhibits MAVS expression by editing its 3'UTR in cells

作　　者：李涛[1] 李京源 朱席琳[1] 伍晓盼[1] 刘英[1] 

机构地区：[1]中国医学科学院基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2017年第6期747-751,共5页Basic and Clinical Medicine

基　　金：国家重点基础研究发展计划(973计划)(2013CB944903)

摘　　要：目的探讨ADAR2在细胞中对线粒体抗病毒信号蛋白MAVS表达的影响及其机制。方法用RT-qRCR检测ADAR家族在细胞中的表达、ADAR2质粒过表达效果以及MAVS的表达;焦磷酸测序验证ADAR2对MAVS的3'UTR区域位点的编辑;双荧光素酶报告质粒检测荧光素酶的相对表达;Western blot检测ADAR2和MAVS蛋白的表达。结果 ADAR家族中ADAR1丰度最高,其次ADAR2也有表达,而ADAR3的表达量很低,几乎检测不到(P<0.05);ADAR2对MAVS的3'UTR区域上的chr20:3869744位点发挥RNA编辑作用(P<0.001);MAVS的3'UTR编辑位点上,编辑型G比正常型A的荧光素酶活性显著降低(P<0.01);在转录和翻译水平,ADAR2都显著降低了MAVS的表达(P<0.05)。结论 ADAR2通过编辑MAVS的3'UTR上的chr20:3869744位点,使其发生A→G的替换,进而抑制了MAVS基因的表达。Objective To investigate the effect of ADAR2 on the expression of MAVS and its mechanism. Methods The RT-qRCR was used to detect the expression of ADAR gene family, the expression level of ADAP,2 and MAVS in cells. The effect of ADAR2 on the 3′UTR region of MAVS was detected by Pyrosequencing. To detect the expression of luciferase by dual luciferase reporter plasmid assay ; The expression of ADAR2 and MAVS were detected by Western blot. Results In the ADAR family, the abundance of ADAR1 was the highest, followed by ADAR2, but the expression of ADAR3 was poor, which was almost impossible to detect（P 〈0. 05）. ADAR2 played a criti- cal role in RNA editing of chr20 ： 3869744 sites on the 3′UTR region of MAVS （ P 〈 0. 001 ）. On the 3′UTR editing site of MAVS, the luciferase activity of the edited G allele was significantly lower than that of the normal A allele （P 〈 0.01 ）. At the level of transcription and translation, ADAR2 significantly inhibited the expression of MAVS （P 〈 0. 05）. Conclusions ADAR2 by editing MAVS＇ 3′UTR on the ehr20：3869744 site, which makes the occurrence of A to G replacement, inhibits the expression of MAVS.

关 键 词：作用于RNA的腺苷脱氨酶2 线粒体抗病毒信号蛋白 RNA编辑 

分 类 号：Q78[生物学—分子生物学]