Chemical tagging for sensitive determination of uridine modifications in RNA.
Chemical tagging for sensitive determination of uridine modifications in RNA.
复制标题
用于灵敏测定 RNA 中尿苷修饰的化学标记
DOI：
10.1039/c9sc05094a
复制
发表时间：
2020-01-08
期刊：
影响因子：
8.4
通讯作者：
Feng YQ
中科院分区：
文献类型：
--
作者：
Cheng QY;Xiong J;Ma CJ;Dai Y;Ding JH;Liu FL;Yuan BF;Feng YQ
信使RNA（mRNA）中动态和可逆修饰的发现为RNA修饰介导的生物过程调控开辟了新的方向。甲基化是mRNA中最常见的修饰，甲基主要修饰在腺嘌呤、胞嘧啶和鸟嘌呤碱基或核糖的2′-羟基上。然而，尿嘧啶碱基（5-甲基尿苷，m5 U）的甲基化尚未在真核生物的mRNA中发现。建立了N-环己基-N′-β-（4-甲基吗啉）乙基碳二亚胺对甲苯磺酸盐（CMCT）标记结合液相色谱-电喷雾电离-质谱（LC-ESI-MS/MS）检测RNA中尿苷修饰的方法。我们的研究结果表明，检测灵敏度的尿苷修饰的RNA增加到1408倍后，CMCT标记。使用开发的方法，我们确定了不同的存在m5 U的mRNA的各种哺乳动物细胞和组织。此外，通过质谱监测的稳定同位素示踪显示，m5 U的甲基来源于S-腺苷-L-甲硫氨酸（SAM）。我们的研究扩展了哺乳动物mRNA中发生的修饰列表。m5 U基因的转录组定位将进一步揭示m5 U在哺乳动物mRNA中的功能作用。信使RNA中动态和可逆修饰的发现为RNA修饰介导的生物过程调控开辟了新的方向。
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, m5U) has not been discovered in mRNA of eukaryotes. In the current study, we established a method of N-cyclohexyl-N′-β-(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 m5U in mRNA of various mammalian cells and tissues. In addition, the stable isotope tracing monitored by mass spectrometry revealed that the methyl group of m5U 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 m5U will further uncover the functional roles of m5U in mRNA of mammals. The discovery of dynamic and reversible modifications in messenger RNA is opening new directions in RNA modification-mediated regulation of biological processes.
登录
查看更多内容
基于金属氧化物的选择性富集结合稳定同位素标记质谱分析核糖缀合物的分析
影响因子：
7.4
作者：
Chu, Jie-Mei;Qi, Chu-Bo;Feng, Yu-Qi
通讯作者：
Feng, Yu-Qi
影响因子：
11.4
作者：
Felden, B;Hanawa, K;Crain, PF
通讯作者：
Crain, PF
化学下拉揭示了哺乳动物转录组的动态假尿苷化
影响因子：
14.8
作者：
Li, Xiaoyu;Zhu, Ping;Yi, Chengqi
通讯作者：
Yi, Chengqi
DOI：
10.1016/j.bbrc.2018.11.104
发表时间：
2019-01-08
影响因子：
3.1
作者：
Chang, Yu-Hsin;Nishimura, Susumu;Takahashi, Satoru
通讯作者：
Takahashi, Satoru
影响因子：
2.9
作者：
HO, NWY;GILHAM, PT
通讯作者：
GILHAM, PT