Bisulfite-sequencing of HBV 1.1 mer-transfected huh7 cells. Bisulfite-sequencing of HBV 1.1 mer-transfected huh7 cells

RNA chemical modifications have been found to play important biological functions. Among which, the 5-methylcytosine (m5C) modification has been reported to participate in viral replication through affecting RNA processing, such as export, decay, translation and so on. In this study, we performed bisulfite sequencing (BS-seq) on HBV 1.1-mer-transfected huh7 cells to identify the m5C sites in HBV mRNA and their function in virus replication was verified. To investigate the mechanism by which m5C methyltransferase NSUN2 suppresses HBV replication, altered global m5C levels in host genes in HBV 1.1-mer-transfected cells were examined by BS-seq. We found that the m5C modification of genes associated with antiviral immunity changed significantly after viral infection. Our study provide new molecular insights into the mechanism of HBV-mediated IFN inhibition Overall design: The changes of m5C modification in huh7 cells after HBV transfection were investigated by bisulfite sequencing.

RNA化学修饰已被证实发挥重要的生物学功能。其中，5-甲基胞嘧啶（5-methylcytosine, m5C）修饰已被报道可通过影响RNA加工过程（包括核输出、降解、翻译等）参与病毒复制调控。本研究对转染乙型肝炎病毒（Hepatitis B Virus, HBV）1.1拷贝型基因组的Huh7细胞进行亚硫酸氢盐测序（bisulfite sequencing, BS-seq），以鉴定HBV mRNA上的m5C位点，并验证其在病毒复制中的功能。为探究m5C甲基转移酶NSUN2抑制HBV复制的分子机制，本研究通过BS-seq检测了HBV 1.1拷贝型基因组转染细胞中宿主基因的整体m5C水平变化。研究发现，抗病毒免疫相关基因的m5C修饰在病毒感染后发生显著改变。本研究为HBV介导的干扰素（IFN）抑制机制提供了全新的分子视角。实验设计概述：本研究通过亚硫酸氢盐测序，探究了HBV转染后Huh7细胞中m5C修饰的变化情况。