SGER: Structure and Function of the SARS Virus RNA Polymerase

SGER：SARS 病毒 RNA 聚合酶的结构和功能

• 批准号: 0341855
• 负责人: C. Cheng Kao
• 金额: $ 10万
• 依托单位: Texas A&M Research Foundation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-10-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0341855&HistoricalAwards=false
• 关键词: SGER; Structure; Function; SARS; Virus

The SARS virus, SCoV, is a newly emerged, highly virulent member of the coronavirus family of positive-strand RNA viruses. While coronaviruses have unique aspects of RNA replication and transcription, little biochemical and structural information exists for the key enzyme of these processes, the RNA-dependent RNA polymerase (RdRp). This SGER project will combine the biochemical and structural biology expertise of the two principal investigators to: 1) use oligonucletoides to construct a cDNA to express the RdRp of the Toronto isolate of the SCoV, purify the protein using conventional and affinity chromatography, establish a polymerase assay, identify the catalytic residues, and map the minimal region required for RNA synthesis in vitro; 2) characterize the initiation of RNA synthesis by the SCoV RdRp; and 3) grow high-resolution crystals of the SCoV RdRp and use X-ray crystallography to elucidate the structure of the protein alone and bound to substrate mimics. Results from these studies will establish biochemical assays and solve the structure for the SCoV polymerase. The results will be highly relevant to the mechanism of viral RNA replication and will allow the design of more effective antivirals against a significant class of human and animal pathogens.

SARS病毒，简称SCOV，是冠状病毒正链RNA病毒家族中一种新出现的高毒力成员。虽然冠状病毒具有RNA复制和转录的独特方面，但这些过程的关键酶-RNA依赖的RNA聚合酶(RdRp)的生化和结构信息很少。这个SGER项目将结合两位主要研究人员的生化和结构生物学专业知识来：1)使用低聚核苷酸构建表达SCOV多伦多分离株RdRp的cDNA，使用常规和亲和层析纯化蛋白质，建立聚合酶分析，鉴定催化残基，并绘制体外合成RNA所需的最小区域图；2)表征SCOV RdRp启动RNA合成的特性；以及3)生长SCOV RdRp的高分辨率晶体，并使用X射线结晶学来阐明蛋白质的结构并与底物模拟物结合。这些研究的结果将建立生化分析并解决SCOV聚合酶的结构。这一结果将与病毒RNA复制的机制高度相关，并将使设计出针对一类重要的人类和动物病原体的更有效的抗病毒药物。