HEPATITIS DELTA VIRUS RNA EDITING

RNA编辑在丁型肝炎的生命周期中发挥着核心作用 病毒，一种亚病毒人类病原体。一种细胞蛋白质，很可能是双重的- 链RNA腺苷脱氨酶，负责编辑 琥珀/W位点的抗基因组RNA。此过程会更改函数 病毒蛋白从一种支持RNA复制到一种 促进病毒粒子的形成和抑制RNA复制。编辑 具有高度的特异性，需要HDV RNA中的特定结构。 这项建议的长期目标是扩大我们的理解 通过腺苷脱氨基进行RNA编辑，以及特定的机制 HDV通过它利用这一过程。转录后 腺苷脱氨的调节作用日益被认为是一种 一种重要的生物调控机制，专门控制 已改变的病毒和细胞蛋白变体的表达 功能。核糖核酸的鉴定与鉴定 所需的结构，负责 可能影响编辑率的修改和辅助因素 和特殊性，仍然是推进我们理解的重要目标 在这一过程中。具体目的是：1)确定RNA的结构 编辑HDV I型RNA所需的；2)识别RNA 编辑HDV基因III RNA所需的结构；3)评估 调控RNA腺苷脱氨酶表达对HDV RNA的影响 编辑、RNA复制、病毒粒子形成和遗传稳定性；以及 4)研究抑制的机制和影响 由唯一的病毒蛋白丁型肝炎抗原编辑。这些目标 将通过多种方法相结合来解决，包括现场- 定向诱变、体外编辑和RNA-蛋白质分析 交互作用，以及对编辑及其结果的评估 转基因细胞。

RNA editing plays a central role in the life cycle of hepatitis delta virus, a subviral human pathogen. A cellular protein, likely a double- stranded RNA adenosine deaminase, is responsible for editing the antigenomic RNA at the amber/W site. This process changes the function of the viral protein from one of supporting RNA replication to one of facilitating virion formation and inhibiting RNA replication. Editing is highly specific, and requires a particular structure in the HDV RNA. The long-term objectives of the proposal are to expand our understanding of RNA editing via adenosine deamination, and the particular mechanisms through which this process is utilized by HDV. Post-transcriptional regulation by adenosine deamination is increasingly recognized as an important biologic regulatory mechanism that specifically controls the expression of viral and cellular protein variants that have altered functions. The identification and characterization of the RNA structures required, the deaminase enzymes responsible for the modifications, and accessory factors that might influence editing rates and specificity, remain important goals in advancing our understanding of this process. The specific aims are 1) to define the RNA structures required for editing HDV genotype I RNA; 2) to identify the RNA structures required for editing HDV genotype III RNA; 3) to evaluate the effects of modulating RNA adenosine deaminase expression on HDV RNA editing, RNA replication, virion formation, and genetic stability; and 4) to examine the mechanisms and implications of the inhibition of editing by hepatitis delta antigen, the sole viral protein. These aims will be addressed by a combination of approaches, including site- directed mutagenesis, analysis in vitro of editing and RNA-protein interactions, and evaluation of editing and its consequences in transfected cells.