ADAR1在转录后水平调控HBV复制的分子机制研究

Hepatitis B virus (HBV) infection poses a serious threat to human health. Host innate immunity factor can resist HBV infection by regulating HBV replication. Being one of the innate immunity factor with a characteristic of deaminase, ADAR1 (Adenosine Deaminase Acting on RNA 1) regulates viral replication through catalyzing hydrolytic deamination of adenosine to inosine on viral RNA. Our preliminary data indicated ADAR1 gene expression increased in liver tissues with HBV infection, and as well as the typical mutation induced by deamination of ADAR1 on RNA in human hepatocellular carcinomas cell transfected with HBV replication plasmid. Our studies reveal that ADAR1 could regulate HBV replication. Our study will focus on the role of ADAR1 plays in HBV replication. We first study the effect of ADAR1 overexpression or silence on HBV replication at DNA and RNA level. Secondly we will investigate the function of deamination of ADAR1 on HBV RNA. Thirdly we will examine the role of ADAR1 on pgRNA encapsidation. Fourthly, we will explore the interaction of ADAR1 and pgRNA and polymerase. Collectively this project will provide a new insight into HBV replication regulation by ADAR1 at the posttranscriptional level and be expected to offer a new strategy for development of anti-viral treatment.

乙型肝炎病毒（HBV）感染是一个严重危及人类健康的公共卫生问题。宿主天然免疫因子可经调控HBV复制抵抗病毒感染。ADAR1为核苷酸脱氨酶类免疫因子，可将病毒RNA特异位点的腺嘌呤脱氨基调控病毒复制。我们研究发现在HBV阳性的小鼠肝组织标本中ADAR1基因表达明显上调；ADAR1过表达后HBV基因X区相应突变增加，提示ADAR1可能参与HBV复制调控。本项目以ADAR1为切入点，以HBV瞬时转染和稳定转染的人肝癌细胞系为模型，评价ADAR1在HBV DNA和RNA水平对HBV复制的影响；明确ADAR1对HBV RNA的脱氨基作用；研究ADAR1对pg RNA包裹的影响；探讨ADAR1与HBV pgRNA及多聚酶蛋白的结合。通过上述研究将可能阐明ADAR1在RNA水平影响HBV复制的分子机制，为寻找抗病毒治疗的靶标奠定基础。

乙型肝炎病毒（HBV）感染严重危害人类健康。宿主天然免疫因子核苷酸脱氨酶如（APOBECs和ADARs）等可以作用于宿主或病毒的DNA或RNA，导致其脱氨基，进而调控病毒复制。RNA编辑酶ADARs在HBV复制过程中作用仍不清楚。本项目通过ADAR1表达特征的研究初步确定了ADAR1和HBV复制的关联，进一步利用Southern blot, Western blot，Realtime PCR,突变分析等技术在不同细胞系（HBV稳定复制肝癌细胞系和瞬时转染肝癌细胞系）中分析研究ADAR1参与HBV复制调控，并初步研究ADAR1调控HBV复制的机制。此研究为寻找病毒复制关键环节为靶点的抗病毒治疗新靶标药物奠定基础，同时本项目也为HBV与宿主细胞相互作用的研究提供新的证据。