中国科技核心期刊
CN:31-1600/Q
ISSN:1004-0374
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《生命科学》 2014, 26(3): 239-247
RNA中的假尿苷修饰:形成、功能及鉴定
李笑雨1#,孙芳芳2#,伊成器1,3*
(1 北京大学生命科学学院,蛋白质与植物基因研究国家重点实验室,北京100871;2 清华大学生命科学学院,北京100084;3 北京大学合成与功能生物分子中心,北京大学-清华大学生命科学联合中心,北京100871)
假尿苷修饰是目前已知丰度最高的RNA 修饰,广泛存在于多个物种的多类RNA 中。作为尿苷的5 位异构体,假尿苷在真核生物中的形成机制主要有两种:依赖于假尿苷合酶或是依赖于H/ACA 核糖核蛋白复合体。假尿苷修饰在多个生物学过程中发挥作用,同时不同位点的假尿苷修饰具有不同的功能。而人为地在mRNA 终止密码子中引入假尿苷修饰则可以使其具有编码能力,这改变了传统的中心法则。目前对于RNA 中假尿苷的研究主要是通过2D-TLC 及液质联用对其进行定量分析,使用CMCT 标记假尿苷及RNase H 和SCARLET 的方法对其进行定位。
 
Pseudouridines in RNA: formation, function and characterization
LI Xiao-Yu1#, SUN Fang-Fang2#, YI Cheng-Qi1,3*
(1 State Key Laboratory of Protein and Plant Gene Research, School of Life Sciences, Peking University, Beijing 100871, China; 2 School of Life Sciences, Tsinghua University, Beijing 100084, China; 3 Peking-Tsinghua Joint Center for Life Science, Synthetic and Functional Biomolecules Center, Peking University, Beijing 100871, China)
Pseudouridine is the most abundant RNA modification and it is widely spread in various RNAs of many organisms. As a rotation isomer of uridine, it is formed mainly through two mechanisms: stand-alone pseudouridine synthases and Box H/ACA Ribonucleoprotein. Pseudouridylation affects many cellular processes, with distinct functions at different sites in RNA. When incorporated into mRNA artifically, nonsense codons-containing pseudouridine can be converted into sense codons. To characterize pseudouridine in RNA, 2D-TLC and LC-MS have been used for quantification purposes, while CMCT-based labeling method, RNase H-based digestion method and most recently “SCARLET” method can all be applied for mapping the pseudouridine sites in RNA.
 
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