Single-Nucleotide Resolution Mapping of <i>N</i><sup>6</sup>-Methyladenine in Genomic DNA
Cheng-Jie Ma, Gaojie Li, Wen-Xuan Shao, Yi-Hao Min, Ping Wang, Jiang-Hui Ding, Neng‐Bin Xie, Min Wang, Feng Tang, Yu‐Qi Feng, Weimin Ci, Yinsheng Wang, Bi‐Feng Yuan
Abstract
High Resolution Image Download MS PowerPoint Slide N 6 -Methyladenine (6mA) is a naturally occurring DNA modification in both prokaryotes and eukaryotes. Herein, we developed a deaminase-mediated sequencing (DM-seq) method for genome-wide mapping of 6mA at single-nucleotide resolution. The method capitalizes on the selective deamination of adenine, but not 6mA, in DNA mediated by an evolved adenine deaminase, ABE8e. By employing this method, we achieved genome-wide mapping of 6mA in Escherichia coli and in mammalian mitochondrial DNA (mtDNA) at single-nucleotide resolution. We found that the 6mA sites are mainly located in the GATC motif in the E. coli genome. We also identified 17 6mA sites in mtDNA of HepG2 cells, where all of the 6mA sites are distributed in the heavy strand of mtDNA. We envision that DM-seq will be a valuable tool for uncovering new functions of 6mA in DNA and for exploring its potential roles in mitochondria-related human diseases.