RNA bisulfite sequencing for polysome fractions

mRNA m5C, which has recently been implicated in the regulation of mRNA mobility, metabolism, and translation, plays important regulatory roles in various biological events. Two types of m5C sites are found in mRNAs. Type I m5C sites, which contain a 3'G-rich triplet motif and locate in the 5' end of hairpin structures, are methylated by NSUN2. Type II m5C sites contain a 3'UCCA motif and locate in the loops of hairpin structures. However, their biogenesis remains unknown. Here we identified a novel mRNA methyltransferase that targets Type II m5C sites and generated BS-seq and RNA-seq data to verify our findings. Overall design: mRNA BS-seq was performed for different polysome fractions of HEK293T cells

信使RNA（mRNA）上的5-甲基胞嘧啶（m5C）修饰近年被证实参与调控mRNA的迁移、代谢与翻译过程，在多种生物学事件中发挥重要调控功能。目前已在mRNA中鉴定出两类m5C修饰位点：第一类m5C位点含有3'端富含G的三联体基序，定位于发夹结构的5'末端，由NSUN2催化甲基化；第二类m5C位点含有3'端UCCA基序，定位于发夹结构的环区。但目前这类位点的生物发生机制仍不明晰。本研究鉴定出一种靶向第二类m5C位点的新型mRNA甲基转移酶，并通过亚硫酸氢盐测序（BS-seq）与RNA测序（RNA-seq）数据验证了该研究发现。实验整体设计：针对HEK293T细胞的不同多聚核糖体组分开展了mRNA BS-seq测序。