Identification and functional characterization of cellular interaction partners of the ribonucleoprotein complex of Borna Disease Virus

博尔纳病病毒核糖核蛋白复合物细胞相互作用伴侣的鉴定和功能表征

• 批准号: 5449165
• 负责人: Professor Dr. Martin Schwemmle
• 金额: --
• 依托单位: Institut für Medizinische Mikrobiologie und Hygiene (IMMH)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2008-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5449165?language=en
• 关键词: Identification; functional; characterization; cellular; interaction

Borna Diesease Virus (BDV) persistently infects the central nervous system and can either cause immunopathology or neurodevelopmental damage in the absence of inflammation. We hypothesize that the BDV replication machinery depends on the interaction with cellular host factors. We further hypothesize that the interaction between viral and cellular factors can trigger neuronal damage. This project therefore focuses on the identification and functional characterization of cellular factors involved in BDV replication and pathogenesis. In a proteomic-based approach, we want to isolate native viral ribonucleoprotein complexes (RNPs), followed by identification of co-purified cellular interactions partners (CIP) using mass spectrometric analysis. Binding studies should further help to define BDV protein mutants that fail to interact with specific CIPs. If these mutants still support BDV minireplicon transcription, we will rescue the corresponding virus mutants, with the help of our newly established reverse genetic system of BDV, to study replication and pathogenesis. In case BDV protein mutants turn out to be non-functional in the minireplicon assay, we will determine whether the corresponding CIP are essential for viral replication. Initially, we will study the role of a known cellular interaction partner of the viral phosphoprotein, HMBG1, a neurite outgrowth factor. Functional characterization of CIP will not only provide important insights into viral replication strategies but also reveal mechanistic details of virus-induced disorders of the CNS.

博尔纳疾病病毒 (BDV) 持续感染中枢神经系统，在没有炎症的情况下可引起免疫病理学或神经发育损伤。我们假设 BDV 复制机制取决于与细胞宿主因子的相互作用。我们进一步假设病毒和细胞因子之间的相互作用可以引发神经元损伤。因此，该项目的重点是 BDV 复制和发病机制中涉及的细胞因子的鉴定和功能表征。在基于蛋白质组学的方法中，我们希望分离天然病毒核糖核蛋白复合物（RNP），然后使用质谱分析鉴定共纯化的细胞相互作用伙伴（CIP）。结合研究应进一步有助于定义无法与特定 CIP 相互作用的 BDV 蛋白突变体。如果这些突变体仍然支持BDV微型复制子转录，我们将借助我们新建立的BDV反向遗传系统来拯救相应的病毒突变体，以研究复制和发病机制。如果 BDV 蛋白突变体在微型复制子测定中被证明没有功能，我们将确定相应的 CIP 是否对病毒复制至关重要。首先，我们将研究病毒磷蛋白的已知细胞相互作用伴侣 HMBG1（一种神经突生长因子）的作用。 CIP 的功能表征不仅将为病毒复制策略提供重要见解，还将揭示病毒引起的中枢神经系统疾病的机制细节。