Detection and quantification of RNA 2′-O-methylation and pseudouridylation

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• 作者：Chao Huang^a ; ^{Chao.Huang@bms.com" class="auth_mail" title="E-mail the corresponding author} ; John Karijolich^b ; Yi-Tao Yu^c ; ^{yitao_yu@urmc.rochester.edu" class="auth_mail" title="E-mail the corresponding author}
• 关键词：Post-transcriptional modification ; RNA ; 2&prime ; -O-methylation ; Pseudouridylation ; Box C/D guide RNA ; Box H/ACA guide RNA
• 刊名：Methods
• 出版年：2016
• 出版时间：1 July 2016
• 年：2016
• 卷：103
• 期：Complete
• 页码：68-76
• 全文大小：1085 K

文摘

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.