当前位置: X-MOL 学术J. Am. Chem. Soc. › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Atomic Mutagenesis of N6-Methyladenosine Reveals Distinct Recognition Modes by Human m6A Reader and Eraser Proteins
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2024-03-06 , DOI: 10.1021/jacs.4c00626
Florian Seitz 1 , Tina Jungnickel 1 , Nicole Kleiber 2 , Jens Kretschmer 2 , Julia Dietzsch 1 , Juliane Adelmann 1 , Katherine E Bohnsack 2 , Markus T Bohnsack 2, 3 , Claudia Höbartner 1, 4
Affiliation  

N6-methyladenosine (m6A) is an important modified nucleoside in cellular RNA associated with multiple cellular processes and is implicated in diseases. The enzymes associated with the dynamic installation and removal of m6A are heavily investigated targets for drug research, which requires detailed knowledge of the recognition modes of m6A by proteins. Here, we use atomic mutagenesis of m6A to systematically investigate the mechanisms of the two human m6A demethylase enzymes FTO and ALKBH5 and the binding modes of YTH reader proteins YTHDF2/DC1/DC2. Atomic mutagenesis refers to atom-specific changes that are introduced by chemical synthesis, such as the replacement of nitrogen by carbon atoms. Synthetic RNA oligonucleotides containing site-specifically incorporated 1-deaza-, 3-deaza-, and 7-deaza-m6A nucleosides were prepared by solid-phase synthesis and their RNA binding and demethylation by recombinant proteins were evaluated. We found distinct differences in substrate recognition and transformation and revealed structural preferences for the enzymatic activity. The deaza m6A analogues introduced in this work will be useful probes for other proteins in m6A research.

中文翻译:


N6-甲基腺苷的原子突变揭示了人类 m6A 阅读器和擦除器蛋白的独特识别模式



N 6 -甲基腺苷 (m 6 A) 是细胞 RNA 中重要的修饰核苷,与多种细胞过程相关,并与疾病有关。与 m 6 A 动态安装和去除相关的酶是药物研究中重点研究的目标,这需要详细了解蛋白质对 m 6 A 的识别模式。在这里,我们利用 m 6 A 的原子诱变系统地研究了两种人类 m 6 A 去甲基化酶 FTO 和 ALKBH5 的机制以及 YTH 阅读器蛋白 YTHDF2/DC1/DC2 的结合模式。原子诱变是指通过化学合成引入的原子特异性变化,例如用碳原子取代氮。通过固相合成制备含有位点特异性掺入的1-脱氮-、3-脱氮-和7-脱氮-m 6 A核苷的合成RNA寡核苷酸,并评估它们的RNA结合和重组蛋白的去甲基化。我们发现底物识别和转化存在明显差异,并揭示了酶活性的结构偏好。这项工作中引入的 deaza m 6 A 类似物将成为 m 6 A 研究中其他蛋白质的有用探针。
更新日期:2024-03-06
down
wechat
bug