Molecular Mechanisms of ZAP/ISG20 mediated HBV RNA Decay

描述（由申请人提供）：干扰素-¿是一种批准的慢性乙型肝炎药物。在筛选干扰素刺激基因（ISG）对HBV复制的潜在抗病毒活性时，我们发现了两种宿主蛋白，特别是锌指抗病毒蛋白（ZAP）和ISG20，它们主要通过转录后下调病毒RNA来抑制HBV复制。我们的数据还表明，ZAP和ISG20都与HBV RNA发生物理相互作用；虽然四个ccch型锌指基序的完整性是zap介导的HBV RNA降解所必需的，但ISG20对HBV RNA的还原取决于其核糖核酸酶活性。有趣的是，酶失活形式的ISG20通过与病毒基因组前RNA （pg）相互作用并随后干扰pgRNA封装来保持对HBV复制的抗病毒活性。此外，ZAP和ISG20在肝细胞中均以基础水平表达，并可在干扰素刺激下进一步诱导表达，表明它们在抑制HBV宿主中的潜在作用。在此，我们建议进一步阐明ZAP/ isg20介导的HBV RNA衰变的分子机制。提出的目标包括：(1)将确定负责ZAP-和isg20介导的RNA降解的HBV RNA序列元件；(2) ZAP和ISG20在先天免疫诱导的抗hbv应答中的生理作用将被确定；(3)将通过蛋白质组学方法鉴定ZAP和ISG20介导的HBV RNA衰变中的辅助因子，并研究ZAP和ISG20之间潜在的合作作用；(4) ZAP和ISG20潜在靶向宿主RNA分子的鉴定，包括mRNA和miRNA。阐明由ZAP和ISG20介导的复杂细胞抗病毒反应的分子机制，将有助于揭示病毒发病过程中的细胞生物学和病毒-宿主相互作用，并有可能导致

DESCRIPTION (provided by applicant): Interferon- ¿ is an approved medication for chronic hepatitis B. In the screening of interferon stimulated genes (ISG) for their potential antiviral activity against HBV replication, we identified two host proteins, specifically zinc finger antivirl protein (ZAP) and ISG20, which inhibit HBV replication primarily through posttranscriptional downregulation of viral RNA. Our data also demonstrated that both ZAP and ISG20 physically interact with HBV RNA; while the integrity of the four CCCH-type zinc finger motifs is required for ZAP-mediated HBV RNA degradation, the reduction of HBV RNA by ISG20 depends on its ribonuclease activity. Interestingly, the enzymatically inactive form of ISG20 retains antiviral activity against HBV replication by interacting with viral pregenomic (pg) RNA and subsequently interfering with pgRNA encapsidation. In addition, both ZAP and ISG20 are expressed at basal levels in hepatocytes, and can be further induced upon interferon stimulation, indicating their potential roles in host restriction of HBV. Here we propose to further elucidate the molecular mechanisms of ZAP/ISG20-mediated HBV RNA decay. The proposed Aims include: (1) The sequence element(s) of HBV RNA responsible for ZAP- and ISG20-mediated RNA degradation will be identified; (2) the physiologic role of ZAP and ISG20 in innate immunity-elicited anti-HBV responses will be determined; (3) co- factor(s) in ZAP- and ISG20-mediated HBV RNA decay will be identified through a proteomic approach, and the potential cooperative role between ZAP and ISG20 will also be investigated; (4) identification of the potential host RNA molecules targeted by ZAP and ISG20, including mRNA and miRNA. Elucidation of the molecular mechanisms of the intricate cellular antiviral response mediated by ZAP and ISG20 will shed light on cell biology and virus-host interaction during viral pathogenesis, and potentially lead to the development of novel therapeutics that exploit host molecules to control viral infection.