Role of Nuclear Interactions of Paramyxoviral Matrix Proteins: Cellular ANP32B in Host Manipulation and Virus Replication

副粘病毒基质蛋白核相互作用的作用：细胞 ANP32B 在宿主操纵和病毒复制中的作用

• 批准号: 280662076
• 负责人: Professor Dr. Stefan Finke
• 金额: --
• 依托单位: Friedrich-Loeffler-Institut - Bundesforschungsinstitut für Tiergesundheit (FLI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2015
• 资助国家: 德国
• 起止时间: 2014-12-31 至 2017-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/280662076?language=en
• 关键词: Role; Nuclear; Interactions; Paramyxoviral; Matrix

We recent identified cellular acidic leucine-rich nuclear phosphoprotein 32 family member B (ANP32B) as a nuclear target for Hendra and Nipah virus matrix protein (M). Further preliminary data show that also another member of the paramyxovirus family is able to interact with ANP32B through M. We therefore conclude that targeting of ANP32B is part of a conserved mechanism, at least within the paramxovirinae subfamily. Identification of the detailed role of this interaction in virus replication and host cell manipulation may lead to identification of a so far unknown mechanism involved in replication and pathogenesis. To clarify the role of ANP32B targeting we here investigate (i) which viral M proteins can interact with the nuclear target, (ii) what sequences in the M proteins are required for specific interaction, (iii) how ANP32B influences virus replication and (iv) whether cellular functions of ANP32B in unconventional nuclear export, gene regulation and apoptosis inhibition are modulated by interacting M proteins. Host cell modulatory activities may play a substantial role in virus replication in infected hosts and thus are of high importance to understand virulence and pathogenesis of viruses. Within the project, established protein interaction assays will be combined with systematic mutagenesis of M proteins and generation of recombinant ANP32B-binding defective virus mutants. Influences on ANP32B functions will be addressed by specific assays for nuclear export, gene regulation and apoptosis detection in the presence of plasmid expressed M proteins and in virus infected cells. In vitro identification of specific M / ANP32B dependent molecular mechanisms and development of binding defective virus mutants will offer the outstanding possibility to investigate the role of ANP32B in disease development and immune escape. Whereas the work program of this three years proposal is focused on in vitro characterization of molecular mechanisms, in continuative work the developed virus mutants and knowledge about affected pathways will allow to investigate the role of the cellular interactor for in vivo replication and pathogenesis of henipa- and other paramyxoviruses. As this project focuses on a host-cell interface that may substantially contribute to replication and host manipulation, the proposed work will not only contribute to a better molecular understanding of virus replication but may also offer novel molecular targets for rational interference with virus replication or disease development.

我们最近确定了细胞酸性富亮氨酸核磷蛋白32家族成员B （ANP32B）作为亨德拉病毒和尼帕病毒基质蛋白(M)的核靶点。进一步的初步数据表明，副粘病毒家族的另一个成员也能够通过m与ANP32B相互作用。因此，我们得出结论，至少在副粘病毒亚家族中，靶向ANP32B是一个保守机制的一部分。鉴定这种相互作用在病毒复制和宿主细胞操作中的详细作用可能导致鉴定迄今未知的参与复制和发病机制的机制。为了阐明ANP32B靶向的作用，我们在这里研究(i)哪些病毒M蛋白可以与核靶标相互作用，（ii） M蛋白中的哪些序列需要特异性相互作用，（iii） ANP32B如何影响病毒复制，以及（iv） ANP32B在非常规核输出、基因调控和细胞凋亡抑制中的细胞功能是否通过相互作用的M蛋白来调节。宿主细胞的调节活动可能在病毒感染宿主的复制过程中发挥重要作用，因此对了解病毒的毒力和发病机制具有重要意义。在该项目中，已建立的蛋白质相互作用分析将与M蛋白的系统诱变和重组anp32b结合缺陷病毒突变体的产生相结合。对ANP32B功能的影响将通过核输出、基因调控和在质粒表达M蛋白和病毒感染细胞中检测细胞凋亡的具体试验来解决。体外鉴定特异性M / ANP32B依赖的分子机制和结合缺陷病毒突变体的发展将为研究ANP32B在疾病发展和免疫逃逸中的作用提供重要的可能性。鉴于这项为期三年的提案的工作计划侧重于体外分子机制的表征，在持续的工作中，开发的病毒突变体和对受影响途径的了解将允许研究细胞相互作用物在henipa和其他副粘病毒的体内复制和发病机制中的作用。由于该项目的重点是宿主-细胞界面，它可能对复制和宿主操作有很大贡献，因此提出的工作不仅有助于更好地了解病毒复制的分子，还可能为合理干扰病毒复制或疾病发展提供新的分子靶点。