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作 者:李晓杰[1] 缪辉[1] 王迪 詹浩淼 王琳琳 吴传芳[1] LI Xiao-Jie;MIAO Hui;WANG Di;ZHAN Hao-Miao;WANG Lin-Lin;WU Chuan-Fang(Department of Functional Genome,College of Life Science,Sichuan University,Chengdu 610064,China)
出 处:《四川大学学报(自然科学版)》2019年第4期743-748,共6页Journal of Sichuan University(Natural Science Edition)
基 金:国家自然科学基金(81490752)
摘 要:为探究RNA与SAFB1蛋白相互作用过程中RNA二级结构的作用.采用足迹法确定SAFB1与LINC-PINT-1特定的结合区域,在此基础上根据RNA结构分析研究SAFB1结合区域所具有特定的二级结构.最后通过凝胶迁移率实验论证该二级结构在RNA与SAFB1蛋白结合中的作用.结果显示:在针对LINC-PINT-1的亚克隆发现只有包含30~55nt区域的亚克隆才能维持RNA结合区域的二级结构,并且证实了LINC-PINT-1与SAFB1蛋白相互作用依赖于特定的RNA二级结构.结果表明:长非编码RNA与SAFB1蛋白相互作用或依赖于特定的RNA二级结构,本研究为RNA与蛋白质相互作用的分子机制研究提供了有力的理论实验支撑.To explore the role of RNA secondary structure in the interaction between RNA and SAFB1 protein,structure analysis and RNA footprinting analysis were adopted.It revealed that SAFB1 binds to a specific region of LINC-PINT-1 and the SAFB1 binding region has a specific secondary structure.In addition,subcloning of LINC-PINT-1 confirmed that subclones contained the 30~55 nt region could maintain this secondary structure.Electrophoretic mobility shift assay(EMSA)found that this secondary structure was involved in the interaction between RNA and SAFB1.In this paper,software prediction and necessary experimental verification have indicated that the interaction between LINC-PINT-1 and SAFB1 depends on the specific RNA secondary structure,which provides a powerful theoretical experimental support for the study of the molecular mechanism of RNA-protein interaction.
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