Structural and Biochemical Characterisation of Bunyavirus L proteins.

布尼亚病毒 L 蛋白的结构和生化特征。

• 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.

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.

布尼亚病毒属于具有负向分段RNA基因组的病毒组。许多新出现的病毒病原体属于布尼亚病毒科。它们能够感染人类和动物，并可能在宿主中引起严重的疾病。由于最近的疫情爆发，新闻中已知的突出例子有汉坦病毒、裂谷热病毒、施马伦贝格病毒和克里米亚-刚果出血热病毒。 这些病毒生命周期的一个关键组成部分是病毒 L 蛋白，这是一种大型多结构域蛋白，含有病毒的 RNA 依赖性 RNA 聚合酶。由于其体积大且结构复杂，有关L蛋白的数据很少。彻底了解这种大分子机器将为医学研究提供重要基础。因此，我们的目标是采用经典生物化学与病毒学和结构生物学相结合的跨学科方法来研究纯化的 L 蛋白。具体目标包括 (1) 在细菌细胞中重组表达和纯化布尼亚病毒 L 蛋白片段，(2) 对这些蛋白进行生化和结构研究，以了解其酶功能并确定潜在的结构同源物。另外（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蛋白以进行功能分析。最后 (4) 我们将使用基于细胞的复制子测定来分析 L 蛋白的特定残基或区域在转录和复制中的作用。这些目标之一的结果将有助于优化其他目标的实验设计。总之，所有结果综合起来将提供 L 蛋白在病毒生命周期中的作用的综合视图。

项目成果

Atomic Structure and Biochemical Characterization of an RNA Endonuclease in the N Terminus of Andes Virus L Protein

安第斯病毒 L 蛋白 N 末端 RNA 核酸内切酶的原子结构和生化特征

• DOI: 10.1371/journal.ppat.1005635
• 发表时间: 2016-06
• 期刊: PLoS Pathogens
• 影响因子: 6.7
• 作者: Fernández-García Y;Reguera J;Busch C;Witte G;Sánchez-Ramos O;Betzel C;Cusack S;Günther S;Reindl S
• 通讯作者: Reindl S

Structural insights into reptarenavirus cap-snatching machinery

爬行病毒抢夺机器的结构见解

• DOI: 10.1371/journal.ppat.1006400
• 发表时间: 2017-05
• 期刊: PLoS Pathogens
• 影响因子: 6.7
• 作者: Rosenthal M;Gogrefe N;Vogel D;Reguera J;Rauschenberger B;Cusack S;Günther S;Reindl S
• 通讯作者: Reindl S

Biochemical characterization of the Lassa virus L protein

拉沙病毒 L 蛋白的生化特征

• DOI: 10.1074/jbc.ra118.006973
• 发表时间: 2019-03-29
• 期刊: The Journal of Biological Chemistry
• 作者: D. Vogel;M. Rosenthal;N. Gogrefe;S. Reindl;S. Günther
• 通讯作者: S. Günther

Biochemical and structural studies reveal differences and commonalities among cap-snatching endonucleases from segmented negative-strand RNA viruses

生化和结构研究揭示分段负链 RNA 病毒抢帽核酸内切酶的差异和共性

• DOI: 10.1074/jbc.ra118.004373
• 发表时间: 2018-12-21
• 期刊: The Journal of Biological Chemistry
• 作者: Holm T;Kopicki JD;Busch C;Olschewski S;Rosenthal M;Uetrecht C;Günther S;Reindl S
• 通讯作者: Reindl S