Single-Base Resolution Mapping Reveals Distinct 5-Formylcytidine in Saccharomyces cerevisiae mRNAs
Single-Base Resolution Mapping Reveals Distinct 5-Formylcytidine in Saccharomyces cerevisiae mRNAs
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单碱基分辨率图谱揭示了酿酒酵母 mRNA 中独特的 5-甲酰胞苷
DOI：
10.1021/acschembio.1c00633
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发表时间：
2022-01-21
影响因子：
4
通讯作者：
Zhou, Xiang
中科院分区：
文献类型：
--
作者：
Wang, Yafen;Chen, Zonggui;Zhou, Xiang
5-甲酰胞苷（Formylcytidine，f（5）C）是一种转录后RNA修饰，位于线粒体tRNA的摆动位置，是线粒体蛋白质合成所必需的。在这里，我们展示了一种检测RNA中f（5）C修饰的方法和一种转录组范围的f（5）C作图技术，称为f（5）C-seq。它是基于吡啶硼烷的处理而开发的，它可以将f（5）C还原为5，6-二氢尿嘧啶，从而在PCR过程中诱导f（5）C位点的C到T转变，实现单碱基分辨检测。通过f（5）C-seq在酿酒酵母中定位后，鉴定了超过1000个f（5）C位点。此外，密码子组成表明在mRNA的摆动位点内偏好f（5）C，这表明在翻译调节中的潜在作用。这些发现扩大了对mRNA中胞嘧啶修饰的理解范围。
5-Formylcytidine (f(5)C) 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 f(5)C modifications in RNA and a transcriptome-wide f(5)C mapping technique, named f(5)C-seq. It is developed based on the treatment of pyridine borane, which can reduce f(5)C to 5,6-dihydrouracil, thus inducing C-to-T transition in f(5)C sites during PCR to achieve single-base resolution detection. More than 1000 f(5)C sites were identified after mapping in Saccharomyces cerevisiae by f(5)C-seq. Moreover, codon composition demonstrated a preference for f(5)C 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.