REGULATION OF ALPHAVIRUS TRANSCRIPTION

描述（改编自申请人摘要）：申请人的长期 目标是开发方法来研究 组装甲病毒复制复合物，转录速率为 监管. 目前，该提案的目标是1。 利用基因分析 探索非结构蛋白在整体复制中的功能， 从负链到正链的转化。 条件 将研究致死突变体以确定nsP 3的作用 RNA复制中的磷酸化，nsP或RNA中的第二位点变化 拯救nsP 2的NTR/解旋酶结构域中的突变体的序列，以及 甲病毒nsP突变体同源干扰的基础。 2. 使用一个 用于研究温度敏感缺陷体外重构系统 在启动子识别、起始、延伸或转化中， 复制复合物（RC-初始）到稳定复制复合物 （RC稳定）的特异性nsP 1 -4突变体。 3. 净化和表征 复制复合物，并确定必要的宿主 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.