College of Chemistry and Molecular Sciences, Wuhan University, Wuhan 430072, Hubei, China.
The Institute of Advanced Studies, College of Life Science, Wuhan University, Wuhan 430072, China.
ACS Chem Biol. 2022 Jan 21;17(1):77-84. doi: 10.1021/acschembio.1c00633. Epub 2021 Nov 30.
5-Formylcytidine (fC) is one type of post-transcriptional RNA modification, which is known at the wobble position of tRNA in mitochondria and essential for mitochondrial protein synthesis. Here, we show a method to detect fC modifications in RNA and a transcriptome-wide fC mapping technique, named fC-seq. It is developed based on the treatment of pyridine borane, which can reduce fC to 5,6-dihydrouracil, thus inducing C-to-T transition in fC sites during PCR to achieve single-base resolution detection. More than 1000 fC sites were identified after mapping in by fC-seq. Moreover, codon composition demonstrated a preference for fC within wobble sites in mRNA, suggesting the potential role in regulation of translation. These findings expand the scope of the understanding of cytosine modifications in mRNA.
5- 甲酰胞苷（fC）是一种转录后 RNA 修饰类型，已知在线粒体 tRNA 的摆动位置，对线粒体蛋白合成至关重要。在这里，我们展示了一种检测 RNA 中 fC 修饰的方法和一种全转录组 fC 作图技术，称为 fC-seq。它是基于吡啶硼烷的处理开发的，吡啶硼烷可以将 fC 还原为 5,6-二氢尿嘧啶，从而在 PCR 过程中诱导 fC 位点的 C 到 T 转换，实现单碱基分辨率检测。通过 fC-seq 在 中进行映射后，鉴定出了 1000 多个 fC 位点。此外，密码子组成在 mRNA 的摆动位置对 fC 表现出偏好，表明其在翻译调控中的潜在作用。这些发现扩展了对 mRNA 中胞嘧啶修饰的理解范围。
