BIOCHEMISTRY OF HEPATITIS C VIRUS REPLICASE

丙型肝炎病毒复制的生物化学

• 批准号: 6382767
• 负责人: Neerja Kaushik-Basu
• 金额: $ 9.07万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-01 至 2004-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6382767
• 关键词: biochemistry; cell free system; enzyme mechanism; enzyme structure; hepatitis C virus; protein protein interaction; protein sequence; replicase; site directed mutagenesis; virus protein

DESCRIPTION (provided by applicant) The long-term goal of the proposed research is to understand and define the biochemical and structural aspects of HCV replicase. This would necessitate identification of the amino acid residues, as well as specific domains on the enzyme molecule involved in the process of copying the viral RNA genome. Recently solved three-dimensional crystal structures of HCV replicase in conjunction with molecular modeling have pinpointed a number of amino acid residues suspected to be involved in specific steps of the enzymatic reaction. These residues will be targeted for site directed mutagenesis and the resulting mutant enzymes will be characterized with respect to their biochemical properties in order to elucidate their functional roles in the catalytic process. Mutagenesis studies along the putative RNA binding track will clarify the roles of these residues in the process of RNA replication. These studies will have direct application in defining the side chain orientation of a number of amino acid residues in the catalytic domain. Knowledge of the complete functional 3-D structure of HCV replicase will help in better understanding of the mechanisms of RNA dependent RNA synthesis and inhibitory action of anti HCV-replicase drugs. Availability of this information, in turn, will further facilitate development of new HCV-replicase directed agents of specific atomic dimensions that would act at specific structural elements of HCV-replicase. The following specific aims are proposed. Aim 1: Biochemical characterization of HCV replicase. Aim 2: To elucidate the catalytic roles of specific amino acid residues in the polymerase function. Aim 3: To determine protein-protein interactions between HCV replicase and other nonstructural viral proteins.

描述（由申请人提供） 拟议研究的长期目标是了解和定义 HCV复制酶的生化和结构方面。这是必要的 鉴定氨基酸残基以及特定域 酶分子参与复制病毒RNA基因组的过程。 最近求解了HCV复制酶的三维晶体结构 与分子建模的结合已确定了许多氨基酸 残基涉嫌参与酶促反应的特定步骤。 这些残基将针对位点定向诱变，并产生 突变酶将以生化为特征 为了阐明其在催化中的功能作用 过程。沿推定RNA结合轨迹的诱变研究将阐明 这些残基在RNA复制过程中的作用。这些研究 将直接应用定义数字的侧链方向 催化域中的氨基酸残基。了解完整 HCV复制酶的功能3-D结构将有助于更好地理解 RNA依赖性RNA合成的机制和抗抑制作用 HCV复制酶药物。此信息的可用性又将进一步 促进特定原子的新型HCV替肽酶定向剂的开发 尺寸将对HCV-复制酶的特定结构元素作用。这 提出了以下特定目标。 目标1：HCV复制酶的生化表征。目标2：阐明 特异性氨基酸残基在聚合酶功能中的催化作用。目的 3：确定HCV复制酶与其他HCV复制酶之间的蛋白质蛋白质相互作用 非结构病毒蛋白。