MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou, China.
College of Animal Sciences, Key Laboratory of Animal Nutrition & Feed Sciences, Ministry of Agriculture, Zhejiang University, Hangzhou, China.
Chem Commun (Camb). 2021 Mar 9;57(20):2499-2502. doi: 10.1039/d0cc08260k.
Here we report a simple and nonradioactive biochemical assay which is capable of accurately determining the substrate methylation sites of human RNA N6-methyladenosine methyltransferases METTL3/METTL14 and METTL16. This method employs enzyme-assisted chemical labelling of a specific base in an RNA substrate with the assistance of an allyl-substituted methyltransferase cofactor, and enables precise identification of the labelling site by a mutation signal from standard nucleic acid sequencing. Our method provides a platform to investigate the enzymatic methylations of long and structurally complex RNA substrates, and facilitates the discovery of new methyltransferases.
在这里，我们报告了一种简单且非放射性的生化分析方法，该方法能够准确地确定人 RNA N6-甲基腺苷甲基转移酶 METTL3/METTL14 和 METTL16 的底物甲基化位点。该方法利用烯丙基取代的甲基转移酶辅助因子辅助在 RNA 底物的特定碱基上进行酶促化学标记，并通过标准核酸测序的突变信号来精确鉴定标记位点。我们的方法为研究长链和结构复杂的 RNA 底物的酶促甲基化提供了一个平台，并有助于发现新的甲基转移酶。
