HEPATITIS DELTA VIRUS RNA EDITING

丁型肝炎病毒 RNA 编辑

• 批准号: 6497092
• 负责人: JOHN L CASEY
• 金额: $ 22.63万
• 依托单位: GEORGETOWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-02-15 至 2003-06-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6497092
• 关键词: RNA binding protein; RNA biosynthesis; adenosine deaminase; gene expression; genetic mapping; hepatitis D; polymerase chain reaction; posttranscriptional RNA processing; site directed mutagenesis; tissue /cell culture; transfection; virus RNA; virus antigen; virus genetics

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.

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，可能是双 链RNA腺苷脱氨酶，负责编辑 琥珀/W 位点的反基因组 RNA。 这个过程改变了函数 病毒蛋白从支持 RNA 复制的一种蛋白转变为支持 RNA 复制的蛋白之一 促进病毒粒子形成并抑制RNA复制。 编辑 具有高度特异性，需要 HDV RNA 具有特定的结构。 该提案的长期目标是扩大我们的理解 通过腺苷脱氨作用进行RNA编辑的研究及其具体机制 HDV 通过该过程来利用该过程。 转录后 腺苷脱氨调节越来越被认为是一种 重要的生物调控机制，专门控制 病毒和细胞蛋白变体的表达已改变 功能。 RNA的鉴定和表征 所需的结构，脱氨酶负责 修改以及可能影响编辑率的辅助因素 和特异性，仍然是增进我们理解的重要目标 这个过程的。 具体目标是 1) 定义 RNA 结构 编辑 HDV 基因型 I RNA 所需； 2) 鉴定RNA 编辑 HDV 基因型 III RNA 所需的结构； 3）评估 调节 RNA 腺苷脱氨酶表达对 HDV RNA 的影响 编辑、RNA 复制、病毒体形成和遗传稳定性；和 4）研究抑制的机制和影响 由丁型肝炎抗原（唯一的病毒蛋白）进行编辑。这些目标 将通过多种方法来解决，包括现场 定向诱变、编辑和 RNA 蛋白的体外分析 相互作用、编辑评价及其后果 转染的细胞。