Structure and function of the ACMV geminivirus replication initiator protein

ACMV双病毒复制起始蛋白的结构和功能

• 批准号: 240618389
• 负责人: Dr. Katharina Hipp
• 金额: --
• 依托单位: Elektronenmikroskopie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/240618389?language=en
• 关键词: Structure; function; ACMV; geminivirus; replication

The replication-associated protein (Rep) of plant geminiviruses is a multifunctional protein necessary for initiation of viral replication by rolling circle replication and for interaction with various host cell factors. In addition, geminivirus Rep proteins possess ATPase/helicase function. African cassava mosaic virus Rep protein will be ectopically expressed in fission yeast (Schizosaccharomyces pombe) and purified from cell extracts for further analysis. By using electron microscopy (EM) and size exclusion chromatography (SEC) the formation of Rep protein oligomers in the presence of different cations as well as adenosine nucleotides and DNA will be assessed. These experiments are aimed to dissect if Rep complexes with different numbers of monomers might be involved for different functions of the Rep protein (e.g. endonucleolytic cleavage for initiation of replication, helicase). Based on these analyses, three dimensional structures of the functional Rep oligomers will be determined by electron microscopy and image processing. The structures will help to understand the different functions that are combined in the Rep protein on a molecular level.

植物双生病毒复制相关蛋白(Rep)是一种多功能蛋白，通过滚环复制启动病毒复制，并与多种寄主细胞因子相互作用。此外，双生病毒Rep蛋白还具有ATPase/Helicase功能。非洲木薯花叶病毒Rep蛋白将在裂殖酵母(Schizosaccharmyces Pombe)中异位表达，并从细胞提取液中纯化以供进一步分析。通过使用电子显微镜(EM)和尺寸排除层析(SEC)，将评估在不同阳离子以及腺苷核苷酸和DNA存在下Rep蛋白寡聚体的形成。这些实验的目的是分析不同数量的单体的Rep复合体是否参与Rep蛋白的不同功能(例如，用于启动复制的内切核裂解、解旋酶)。在这些分析的基础上，将通过电子显微镜和图像处理来确定功能性Rep低聚物的三维结构。这些结构将有助于在分子水平上理解Rep蛋白中结合的不同功能。

项目成果

The RXL motif of the African cassava mosaic virus Rep protein is necessary for rereplication of yeast DNA and viral infection in plants.

• DOI: 10.1016/j.virol.2014.06.002
• 发表时间: 2014-08
• 期刊: Virology
• 影响因子: 3.7
• 作者: K. Hipp;P. Rau;Benjamin Schäfer;B. Gronenborn;H. Jeske