Single-base resolution mapping of 2′-O-methylation sites by an exoribonuclease-enriched chemical method  

作　　者：Ping Zhang Junhong Huang Wujian Zheng Lifan Chen Shurong Liu Anrui Liu Jiayi Ye Jie Zhou Zhirong Chen Qiaojuan Huang Shun Liu Keren Zhou Lianghu Qu Bin Li Jianhua Yang 

机构地区：[1]MOE Key Laboratory of Gene Function and Regulation,State Key Laboratory of Biocontrol,School of Life Sciences,The Fifth Affiliated Hospital,Sun Yat-sen University,Guangzhou 510275,China [2]Department of Chemistry,The University of Chicago,Chicago,IL 60637,USA [3]Department of Systems Biology,Beckman Research Institute of City of Hope,Monrovia,CA 91016,USA

出　　处：《Science China(Life Sciences)》2023年第4期800-818,共19页中国科学（生命科学英文版）

基　　金：supported by the National Key R&D Program of China(2019YFA0802202);the National Natural Science Foundation of China(91940304,31971228,31900903,31970604,32100467,32225011);the Youth Science and Technology Innovation Talent of Guangdong Te Zhi Plan(2019TQ05Y181)。

摘　　要：2′-O-methylation(Nm)is one of the most abundant RNA epigenetic modifications and plays a vital role in the post-transcriptional regulation of gene expression.Current Nm mapping approaches are normally limited to highly abundant RNAs and have significant technical hurdles in m RNAs or relatively rare non-coding RNAs(nc RNAs).Here,we developed a new method for enriching Nm sites by using RNA exoribonuclease and periodate oxidation reactivity to eliminate 2′-hydroxylated(2′-OH)nucleosides,coupled with sequencing(Nm-REP-seq).We revealed several novel classes of Nm-containing nc RNAs as well as m RNAs in humans,mice,and drosophila.We found that some novel Nm sites are present at fixed positions in different t RNAs and are potential substrates of fibrillarin(FBL)methyltransferase mediated by sno RNAs.Importantly,we discovered,for the first time,that Nm located at the 3′-end of various types of nc RNAs and fragments derived from them.Our approach precisely redefines the genome-wide distribution of Nm and provides new technologies for functional studies of Nm-mediated gene regulation.

关 键 词：2′-O-methylation MgR exoribonuclease periodate oxidation epitranscriptome TRANSCRIPTOME 

分 类 号：Q75[生物学—分子生物学]