Next-Generation Sequencing-Based Analysis of the Effects of N1- and N6-Methyldeoxyadenosine Adducts on DNA Transcription,Analytical Chemistry

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Next-Generation Sequencing-Based Analysis of the Effects of N1- and N6-Methyldeoxyadenosine Adducts on DNA Transcription
Analytical Chemistry ( IF 6.7 ) Pub Date : 2022-08-03 , DOI: 10.1021/acs.analchem.2c01764
Ying Yang ₁ , Ziyu Wang ₁ , Juan Wang ₁ , Xiaoxia Dai ₁ , Changjun You ₁

Affiliation

DNA methylation can occur naturally or be induced by various environmental and chemotherapeutic agents. The regioisomeric N¹- and N⁶-methyldeoxyadenosine (1mdA and 6mdA, respectively) represent an important class of methylated DNA adducts. In this study, we developed a shuttle vector- and next-generation sequencing-based assay to quantitatively assess the effects of 1mdA and 6mdA on the accuracy and efficiency of DNA transcription. Our results revealed that 1mdA can induce multiple types of mutant transcripts and strongly inhibit DNA transcription, whereas 6mdA is a nonmutagenic DNA adduct that can exhibit a subtle but significant inhibitory effect on DNA transcription in vitro and in human cells. Moreover, our results demonstrated that the transcription-coupled nucleotide excision repair pathway is dispensable for the removal of 1mdA and 6mdA from the template DNA strand in human cells. These findings provided new important insights into the functional interplay between DNA methylation modifications and transcription in mammalian cells.

中文翻译：

N1-和N6-甲基脱氧腺苷加合物对DNA转录影响的下一代测序分析

DNA甲基化可以自然发生或由各种环境和化学治疗剂诱导。区域异构N ¹ - 和N ⁶-甲基脱氧腺苷（分别为 1mdA 和 6mdA）代表一类重要的甲基化 DNA 加合物。在这项研究中，我们开发了一种基于穿梭载体和下一代测序的检测方法，以定量评估 1mdA 和 6mdA 对 DNA 转录准确性和效率的影响。我们的研究结果表明，1mdA 可以诱导多种类型的突变转录物并强烈抑制 DNA 转录，而 6mdA 是一种非诱变 DNA 加合物，可以在体外和人体细胞中对 DNA 转录表现出微妙但显着的抑制作用。此外，我们的结果表明，转录偶联的核苷酸切除修复途径对于从人类细胞的模板 DNA 链中去除 1mdA 和 6mdA 是可有可无的。

更新日期：2022-08-03

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