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.

流感病毒（IAVS）负责复发性流行流行病和偶尔发生毁灭性的大流行病。它们的分割基因组由8个VRNA组成，有利于进化，但需要一种复杂的包装机制，仍然知之甚少。 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，有自己的代码。我们的目的是在VRNA和NP级别上解码VRNP相互作用网络。为此，我们将将VRNP的结构探测与几个化学探针和RNA-RNA在野生型和定义的突变病毒内部颗粒内部的交联以及由于VRNA或NP突变引起的包装决定因素的改变。我们的项目通过结合三个关键方面来扩展以前的研究。首先，通过对野生型和突变病毒进行结构分析，我们将克服难以识别VRNA-VRNA和VRNA-NP相互作用对于包装IAV基因组至关重要的IAV基因组至关重要的IAV基因组。其次，通过完整和拆卸的病毒颗粒获得的RNA探测和RNA交联数据的系统比较将使相邻VRNP之间的相互作用明确鉴定。第三，通过对几种与RNA不同部分反应的试剂进行RNA探测，我们将能够区分VRNA-VRNA相互作用与VRNA-NP相互作用。总的来说，我们的项目应提供原理证明，表明特定的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，并为这些相互作用的可塑性提供了结构灯。