Chemical tagging for sensitive determination of uridine modifications in RNA

被引：43

作者：

Cheng, Qing-Yun ^{[1
]}

Xiong, Jun ^{[1
]}

Ma, Cheng-Jie ^{[1
]}

Dai, Yi ^{[1
]}

Ding, Jiang-Hui ^{[1
]}

Liu, Fei-Long ^{[1
]}

Yuan, Bi-Feng ^{[1
]}

Feng, Yu-Qi ^{[1
]}

机构：

[1] Wuhan Univ, Key Lab Analyt Chem Biol & Med, Minist Educ, Sauvage Ctr Mol Sci,Dept Chem, Wuhan 430072, Peoples R China

来源：

CHEMICAL SCIENCE | 2020年 / 11卷 / 07期

基金：

中国国家自然科学基金; 国家重点研发计划;

关键词：

MESSENGER-RNA; GENE-EXPRESSION; MASS-SPECTROMETRY; CHARGE-REMOTE; SEQUENCING TECHNOLOGIES; SELECTIVE ENRICHMENT; MODIFIED NUCLEOTIDES; DNA METHYLATION; NUCLEIC-ACIDS; REVEALS;

D O I：

10.1039/c9sc05094a

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

The discovery of dynamic and reversible modifications in messenger RNA (mRNA) is opening new directions in RNA modification-mediated regulation of biological processes. Methylation is the most prevalent modification occurring in mRNA and the methyl group is mainly decorated in the adenine, cytosine, and guanine base or in the 2 '-hydroxyl group of ribose. However, methylation of the uracil base (5-methyluridine, m(5)U) has not been discovered in mRNA of eukaryotes. In the current study, we established a method of N-cyclohexyl-N '-beta-(4-methylmorpholinium) ethylcarbodiimide p-toluenesulfonate (CMCT) labelling coupled with liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS/MS) analysis for the sensitive determination of uridine modifications in RNA. Our results demonstrated that the detection sensitivities of uridine modifications in RNA increased up to 1408 fold upon CMCT labelling. Using the developed method, we identified the distinct existence of m(5)U in mRNA of various mammalian cells and tissues. In addition, the stable isotope tracing monitored by mass spectrometry revealed that the methyl group of m(5)U originated from S-adenosyl-l-methionine (SAM). Our study expanded the list of modifications occurring in mRNA of mammals. Future work on transcriptome-wide mapping of m(5)U will further uncover the functional roles of m(5)U in mRNA of mammals.

引用

页码：1878 / 1891

页数：14

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