Detection and quantification of RNA 2'-O-methylation and pseudouridylation.
Detection and quantification of RNA 2'-O-methylation and pseudouridylation.
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DOI：
10.1016/j.ymeth.2016.02.003
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发表时间：
2016-07-01
期刊：
影响因子：
--
通讯作者：
Yu YT
中科院分区：
文献类型：
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作者：
Huang C;Karijolich J;Yu YT
RNA介导的RNA修饰是一种自然发生的过程，它将2 ' - o甲基化和假尿嘧啶化引入rRNA、剪接体snRNA和其他几种类型的RNA中。Box C/D核糖核蛋白（RNP）和Box H/ACA核糖核蛋白（RNP）分别含有一种独特的引导RNA （Box C/D RNA或Box H/ACA RNA）和一组核心蛋白，分别负责2 ' - o -甲基化和假尿嘧啶化。框C/D RNA和框H/ACA RNA通过碱基配对与其RNA底物提供修饰特异性。这些转录后修饰可以深刻地改变底物rna的性质和功能。因此，需要建立可靠的、标准化的修饰方法来研究修饰后的rna核苷酸的生物学功能。在这里，我们提出了几种检测和定量转录后2 ' - o甲基化和假尿嘧啶化的敏感和有效的方法和方案。
RNA-guided RNA modification is a naturally occurring process that introduces 2′-O-methylation and pseudouridylation into rRNA, spliceosomal snRNA and several other types of RNA. The Box C/D ribonucleoproteins (RNP) and Box H/ACA RNP, each containing one unique guide RNA (Box C/D RNA or Box H/ACA RNA) and a set of core proteins, are responsible for 2′-O-methylation and pseudouridylation respectively. Box C/D RNA and Box H/ACA RNA provide the modification specificity through base pairing with their RNA substrate. These post-transcriptional modifications could profoundly alter the properties and functions of substrate RNAs. Thus it is desirable to establish reliable and standardized modification methods to study biological functions of modified nucleotides in RNAs. Here, we present several sensitive and efficient methods and protocols for detecting and quantifying post-transcriptional 2′-O-methylation and pseudouridylation.