Structural and Biochemical Characterisation of Bunyavirus L proteins.

布尼亚病毒 L 蛋白的结构和生化特征。

基本信息

批准号：
262051160
负责人：
Dr. Sophia Reindl
金额：
--
依托单位：
Abteilung Virologie
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2014
资助国家：
德国
起止时间：
2013-12-31 至 2017-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/262051160?language=en
关键词：
Structural Biochemical Characterisation Bunyavirus proteins.

项目摘要

Bunyaviruses belong to the group of viruses that possess a segmented RNA genome in negative orientation. Many of the newly emerged viral pathogens belong to the family of Bunyaviridae. They are capable of infecting both humans and animals and can cause severe diseases in their hosts. Prominent examples known from the news due to recent outbreaks are Hantaviruses, the Rift-Valley Fever Virus, the Schmallenberg Virus and the Crimean-Congo-Hemorrhagic Fever Virus. A key component in the lifecycle of these viruses is the viral L protein, a large multi-domain protein containing the RNA-dependant RNA polymerase of the virus. Due to its large size and complex structure, data on the L protein is scarce. A thorough understanding of this large molecular machine will provide an important basis for medical research. We therefore aim at studying the purified L protein using a interdisciplinary approach combining classical biochemistry with virology and structural biology. Specific objectives include (1) the recombinant expression and purification of fragments of the Bunyavirus L protein in bacterial cells, (2) the biochemical and structural investigation of these proteins to understand their enzymatic function and determine potential structural homologs. In addition (3) we will purify full-length L protein using insect cell or mammalian cell expression systems for functional analyses. Finally (4) we will use cell based replicon assays to analyse the role of specific residues or regions of the L protein in transcription and replication. Results from one of these objectives will aid in optimization of the experimental design for the other objectives. In summary, all results taken together will provide an integrated view of the role of the L protein in the viral life cycle.

Bunyavirus属于具有分段RNA基因组为阴性取向的病毒组。许多新出现的病毒病原体属于Bunyaviridae家族。他们能够感染人类和动物，并可能在宿主中引起严重的疾病。由于最近爆发而从新闻中知道的突出例子是汉坦病毒，裂谷发烧病毒，施马伦贝格病毒和克里米亚 - 隆戈 - 雷尔毛发病毒。这些病毒生命周期中的关键成分是病毒L蛋白，该病毒蛋白是一种大型多域蛋白，含有该病毒的RNA依赖性RNA聚合酶。由于其尺寸较大和复杂的结构，因此L蛋白上的数据很少。对这种大分子机器的透彻理解将为医学研究提供重要的基础。因此，我们旨在使用将经典生物化学与病毒学和结构生物学结合的跨学科方法研究纯化的L蛋白。具体目标包括（1）细菌细胞中Bunyavirus L蛋白片段的重组表达和纯化，（2）这些蛋白质的生化和结构研究，以了解其酶促功能并确定潜在的结构同源物。此外，（3）我们将使用昆虫细胞或哺乳动物细胞表达系统纯化全长L蛋白进行功能分析。最后（4）我们将使用基于细胞的复制子分析来分析L蛋白在转录和复制中的特定残基或区域的作用。这些目标之一的结果将有助于优化其他目标的实验设计。总而言之，所有结果将共同提供对L蛋白在病毒生命周期中的作用的综合视图。