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。我们推测BDV的复制机制依赖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，本项目的重点是参与BDV复制和发病机制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伴侣（CIP）。结合研究应进一步帮助确定BDV蛋白突变体，不能与特定的CIP相互作用。如果这些突变体仍然支持BDV小复制子的转录，我们将拯救相应的病毒突变体，在我们新建立的BDV反向遗传系统的帮助下，研究复制和致病性。如果BDV蛋白突变体在微复制子测定中被证明是无功能的，我们将确定相应的CIP是否是病毒复制所必需的。首先，我们将研究一个已知的病毒磷蛋白，HMBG1，神经突生长因子的细胞相互作用的合作伙伴的作用。CIP的功能表征不仅将提供重要的见解病毒复制策略，但也揭示了病毒引起的中枢神经系统疾病的机制细节。