核糖核酸
RNA甲基化
5-甲基胞嘧啶
核糖体RNA
化学
5.8S核糖体RNA
RNA结合蛋白
分子生物学
DNA
生物
5S核糖体RNA
细胞生物学
基因
非编码RNA
生物化学
DNA甲基化
甲基化
甲基转移酶
基因表达
核糖体
作者
Xiaoxia Dai,Gwendolyn González,Lin Li,Jie Li,Changjun You,Weili Miao,Jiawei Hu,Lijuan Fu,Yue Zhao,Ruidong Li,Lichao Li,Xuemei Chen,Yanhui Xu,Weifeng Gu,Yinsheng Wang
标识
DOI:10.1021/acs.analchem.9b04505
摘要
5-Methylcytosine is found in both DNA and RNA; although its functions in DNA are well established, the exact role of 5-methylcytidine (m5C) in RNA remains poorly defined. Here we identified, by employing a quantitative proteomics method, multiple candidate recognition proteins of m5C in RNA, including several YTH domain-containing family (YTHDF) proteins. We showed that YTHDF2 could bind directly to m5C in RNA, albeit at a lower affinity than that toward N6-methyladenosine (m6A) in RNA, and this binding involves Trp432, a conserved residue located in the hydrophobic pocket of YTHDF2 that is also required for m6A recognition. RNA bisulfite sequencing results revealed that, after CRISPR-Cas9-mediated knockout of the YTHDF2 gene, the majority of m5C sites in rRNA (rRNA) exhibited substantially augmented levels of methylation. Moreover, we found that YTHDF2 is involved in pre-rRNA processing in cells. Together, our data expanded the functions of the YTHDF2 protein in post-transcriptional regulations of RNA and provided novel insights into the functions of m5C in RNA biology.
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