Ma Cheng-Jie, Li Gaojie, Shao Wen-Xuan, Min Yi-Hao, Wang Ping, Ding Jiang-Hui, Xie Neng-Bin, Wang Min, Tang Feng, Feng Yu-Qi, Ci Weimin, Wang Yinsheng, Yuan Bi-Feng
School of Public Health, Department of Radiation and Medical Oncology, Zhongnan Hospital of Wuhan University, Wuhan University, Wuhan 430071, China.
Sauvage Center for Molecular Sciences, Department of Chemistry, Wuhan University, Wuhan 430072, China.
ACS Cent Sci. 2023 Aug 28;9(9):1799-1809. doi: 10.1021/acscentsci.3c00481. eCollection 2023 Sep 27.
-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 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 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.
N6-甲基腺嘌呤（6mA）是原核生物和真核生物中自然存在的一种DNA修饰。在此，我们开发了一种脱氨酶介导的测序（DM-seq）方法，用于在单核苷酸分辨率下对全基因组范围的6mA进行定位。该方法利用了一种经过进化的腺嘌呤脱氨酶ABE8e介导的DNA中腺嘌呤而非6mA的选择性脱氨作用。通过采用这种方法，我们在细菌基因组和哺乳动物线粒体DNA（mtDNA）中实现了单核苷酸分辨率下全基因组范围的6mA定位。我们发现，细菌基因组中的6mA位点主要位于GATC基序中。我们还在HepG2细胞的mtDNA中鉴定出17个6mA位点，其中所有的6mA位点都分布在mtDNA的重链上。我们设想，DM-seq将成为揭示DNA中6mA新功能以及探索其在与线粒体相关的人类疾病中的潜在作用的有价值工具。
