Decoding the vRNP interaction network of influenza A viruses required for genome packaging

解码基因组包装所需的甲型流感病毒的 vRNP 相互作用网络

• 批准号: 431323641
• 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: 2019
• 资助国家: 德国
• 起止时间: 2018-12-31 至 2022-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/431323641?language=en
• 关键词: Decoding; vRNP; interaction; network; influenza

Influenza A viruses (IAVs) are responsible for recurrent flu epidemics and occasional devastating pandemics. Their segmented genome consisting of 8 vRNAs favors evolution but requires a complex packaging mechanism that remains poorly understood. Our work and work by others indicate that the segmented genome of IAVs build a supramolecular complex held together by a redundant vRNA-vRNA interaction network and reveals a complex interplay between the vRNA packaging signals and the viral nucleoprotein (NP), suggesting that each viral ribonucleoprotein (vRNP), formed by the association of a vRNA with a polymerase complex and multiple copies of NP, has its own code. Our aim is to decode the vRNP interaction network at the vRNA and NP levels. To that goal, we will combine structural probing of vRNPs with several chemical probes and RNA-RNA crosslinking inside particles of wild type and defined mutant viruses with altered packaging determinants due to mutations in vRNAs or NP. Our project extends previous studies by combining three crucial aspects. First, by performing our structural analysis on wild type and mutant viruses, we will overcome the difficulty of identifying vRNA-vRNA and vRNA-NP interactions crucial for packaging of the IAV genome that is inherent to the redundancy of these interactions. Second, the systematic comparison of the RNA probing and RNA crosslinking data obtained with intact and disassembled viral particles will allow the unambiguous identification of interactions between adjacent vRNPs. Third, by performing RNA probing with several reagents that react with different parts of RNA, we will be able to discriminate vRNA-vRNA interactions from vRNA-NP interactions. Altogether, the our project should provide the proof of principle that specific RNA-RNA and/or RNA-protein interactions are essential for the IAV genome packaging process and shed a structural light on the plasticity of these interactions.

甲型流感病毒 (IAV) 是造成反复流行的流感和偶尔发生的毁灭性流行病的原因。它们的分段基因组由 8 个 vRNA 组成，有利于进化，但需要复杂的包装机制，但人们对此仍知之甚少。我们的工作和其他人的工作表明，IAV 的分段基因组构建了一个由冗余 vRNA-vRNA 相互作用网络结合在一起的超分子复合物，并揭示了 vRNA 包装信号和病毒核蛋白 (NP) 之间复杂的相互作用，这表明每种病毒核糖核蛋白（vRNP）由 vRNA 与聚合酶复合物和 NP 的多个拷贝结合而成，有自己的代码。我们的目标是在 vRNA 和 NP 水平上解码 vRNP 相互作用网络。为了实现这一目标，我们将对 vRNP 的结构探测与多种化学探针以及野生型和特定突变病毒颗粒内的 RNA-RNA 交联结合起来，这些突变病毒由于 vRNA 或 NP 的突变而改变了包装决定因素。我们的项目通过结合三个关键方面来扩展之前的研究。首先，通过对野生型和突变病毒进行结构分析，我们将克服识别 vRNA-vRNA 和 vRNA-NP 相互作用的困难，这些相互作用对于 IAV 基因组的包装至关重要，而这些相互作用的冗余是固有的。其次，通过完整和分解的病毒颗粒获得的 RNA 探测和 RNA 交联数据的系统比较将允许明确识别相邻 vRNP 之间的相互作用。第三，通过使用与 RNA 不同部分反应的几种试剂进行 RNA 探测，我们将能够区分 vRNA-vRNA 相互作用和 vRNA-NP 相互作用。总而言之，我们的项目应该提供原理证明，证明特定的 RNA-RNA 和/或 RNA-蛋白质相互作用对于 IAV 基因组包装过程至关重要，并从结构上阐明这些相互作用的可塑性。