REGULATION OF ALPHAVIRUS TRANSCRIPTION

描述(改编自申请者摘要)：申请者的长距离 目标是开发方法来研究 组装了甲型病毒复制复合体，转录速度为 受监管的。她在目前的提案中的目标是1.使用基因分析 探讨非结构蛋白在整体复制和表达中的作用 在从负链到正链合成的转换形式中。有条件的 将对致死突变进行研究以确定NSP3的作用 RNA复制中的磷酸化，即NSP或RNA的第二位点变化 在nsP2的NTPase/解旋酶结构域中拯救突变的序列，以及 甲型病毒NSP突变体同源干扰的基础。2.要使用 用于研究温度敏感缺陷的体外重建系统 在启动子的识别、起始、延长或转化中 复制复合体(RC-初始)到稳定的复制复合体 (RC-稳定)的特定nsP1-4突变体。3.提纯和表征 从受感染细胞复制复合体并识别必要宿主 各种因素。

DESCRIPTION (Adapted from Applicant's Abstract): The applicant's long range goal is to develop methods to investigate the mechanism by which the alphavirus replication complex is assembled and the rate of transcription is regulated. Her aims in the current proposal are 1. To use genetic analysis to probe the functions of nonstructural proteins in overall replication and in the conversion form minus strand to plus strand synthesis. Conditional lethal mutants will be studied to identify the role of the nsP3 phosphorylation in RNA replication, the second-site changes in nsP or RNA sequences rescuing mutants in the NTPase/helicase domain of nsP2, and the basis of homologous interference by alphavirus nsP mutants. 2. To use an in vitro reconstitution system to investigate temperature sensitive defects in promoter recognition, initiation, elongation or conversion of initial replication complexes (RC-initial) to stable replication complexes (RC-stable) of specific nsP1-4 mutants. 3. To purify and characterize replication complexes from infected cells and to identify essential host factors.