BIOCHEMISTRY OF HEPATITIS C VIRUS REPLICASE

描述(由申请人提供) 拟议研究的长期目标是理解和定义 丙型肝炎病毒复制酶的生化和结构方面。这将是必要的 氨基酸残基的鉴定，以及特定的结构域 参与病毒RNA基因组复制过程的酶分子。 最近解开了丙型肝炎病毒复制酶的三维晶体结构 与分子建模相结合已经精确地确定了一些氨基酸 怀疑与酶反应的特定步骤有关的残基。 这些残基将被定位为定点突变和由此产生的 突变的酶将根据它们的生化特性进行表征 性质，以阐明它们在催化中的功能作用 进程。沿假定的RNA结合轨迹的突变研究将澄清 这些残基在RNA复制过程中的作用。这些研究 将直接应用于定义数字的侧链方向 催化区的氨基酸残基。完整的知识 丙型肝炎病毒复制酶的功能三维结构将有助于更好地理解 RNA依赖的RNA合成机制及抗肿瘤药物的抑制作用 丙型肝炎病毒复制酶药物。这些信息的可获得性反过来又将进一步 促进新的丙型肝炎病毒复制酶导向的特异性原子试剂的开发 这些维度将作用于丙型肝炎病毒复制酶的特定结构元素。这个 提出了以下具体目标。 目的1：丙型肝炎病毒复制酶的生化特性。目标2：阐明 特定氨基酸残基在聚合酶功能中的催化作用。目标 3：确定丙型肝炎病毒复制酶和其他蛋白之间的蛋白质相互作用 非结构病毒蛋白。

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.