MECHANISTIC STUDIES ON SINDBIS VIRUS RNA REPLICATION

辛德毕斯病毒是甲病毒家族的一员，该家族包含许多人类和动物病原体。 该提案的目标是在分子水平上了解辛德毕斯病毒负链RNA合成启动所涉及的事件。 体外复制系统将用于检查辛德毕斯病毒负链RNA合成的最低顺式作用要求。 使用代表负链RNA合成诱变的最小模板的小模板RNA，将使用模板竞争和交联试验来确定模板识别的要求。使用RNA合成抑制剂的生化分析旨在确定起始和延长复制酶复合物的稳定性。 最后，使用各种模板RNA，将确定除最小启动子以外的序列在负链合成中的作用。 这些研究将扩展到正链RNA合成。

Sindbis virus is a member of the Alphavirus family which contains a number of human and animal pathogens. The goal of the proposal is to obtain an understanding at the molecular level of the events involved in the initiation of Sindbis virus minus strand RNA synthesis. An in vitro replication system will be used to examine the minimum cis-acting requirements for Sindbis virus minus-strand RNA synthesis. Using small template RNAs representing the minimal template for minus strand RNA synthesis mutagenesis, template competition and crosslinking assays will be used to determine the requirements for template recognition. Biochemical analyses using inhibitors of RNA synthesis will be aimed at determining the stability of the initiating and elongating replicase complexes. Finally using various template RNAs the role of sequences other than the minimal promoter in minus strand synthesis will be determined. These studies will be extended to plus-strand RNA synthesis.