A Mutation‐Based Method for Pinpointing a DNA N6‐Methyladenine Methyltransferase Modification Site at Single Base Resolution. (22nd April 2021)

• Record Type: Journal Article
• Authors: Cheng, Mohan; Shu, Xiao; Cao, Jie; Gao, Minsong; Xiang, Siying; Wang, Fengqin; Wang, Yizhen; Liu, Jianzhao
• Is Part Of: Chembiochem. Volume 22:Number 11(2021)
• Page Start: 1936
• Page End: 1939
• Publication Date: 2021-04-22
• Subjects: DNA methylation -- Detection of DNA 6mA at single base resolutionDNA methyltransferase-assisted chemical labeling -- DNA replication -- Engineered methyltransferase cofactor; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202100088
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: DNA N 6 ‐methyladenine (6mA) has recently received notable attention due to an increased finding of its functional roles in higher eukaryotes. Here we report an enzyme‐assisted chemical labeling method to pinpoint the DNA 6mA methyltransferase (MTase) substrate modification site at single base resolution. A designed allyl‐substituted MTase cofactor was applied in the catalytic transfer reaction, and the allyl group was installed to the N 6 ‐position of adenine within a specific DNA sequence to form N 6 ‐allyladenine (6aA). The iodination of 6aA allyl group induced the formation of 1, N 6 ‐cyclized adenine which caused mutations during DNA replication by a polymerase. Thus the modification site could be precisely detected by a mutation signal. We synthesized 6aA deoxynucleoside and deoxynucleotide model compounds and a 6aA‐containing DNA probe, and screened nine DNA polymerases to define an optimal system capable of detecting the substrate modification site of a DNA 6mA MTase at single‐base resolution. Abstract : An enzyme‐assisted chemical labeling method to pinpoint the DNA 6mA methyltransferase (MTase) substrate modification site at single‐base resolution is described.