Examining the Dynamic Architecture of the Influenza Virus Genome

检查流感病毒基因组的动态结构

基本信息

批准号：
10282637
负责人：
Nara Lee
金额：
$ 19.56万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-06-01 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10282637
关键词：
Address Architecture Area Binding Cell Nucleus Cell membrane Cells Cessation of life Complex Cytoplasm Data Development Endosomes Epidemic Genome High-Throughput Nucleotide Sequencing Immunoprecipitation In Vitro Individual Infection Influenza Influenza A virus Integration Host Factors Knowledge Life Cycle Stages Ligation Light Link Maps Modeling Nuclear Nucleoproteins Nucleotides Outcome Phase Polymerase Positioning Attribute Process RNA RNA Binding RNA Transport RNA chemical synthesis RNA-Binding Proteins Resolution Ribonucleoproteins Route Site Specificity Structure Surveillance Program Techniques Time Travel Viral Viral Genome Virion Virus Assembly Virus Replication crosslink crosslinking and immunoprecipitation sequencing design experimental study genome-wide influenzavirus insight next generation sequencing pandemic disease particle pathogen spatiotemporal viral RNA viral fitness whole genome

项目摘要

Assembly of influenza viruses is a complex coordinated process. The influenza virus genome consists of eight negative-sense single-stranded RNA segments that are coated by virally encoded nucleoprotein (NP) oligomers. During infection, the newly synthesized viral RNA (vRNA) and NP associate in the nucleus. The viral ribonucleoprotein (vRNP) segments, composed of vRNA, NP and the viral polymerase complex, are transported to the plasma membrane, where all eight vRNP segments are packaged into a single virion. The interaction between vRNA and NP not only serves to protect vRNA inside the host cell, but also promotes transport of the vRNP segments into and out of the nucleus, vRNA synthesis, and selective packaging of all eight vRNA segments. We have recently shown using techniques entailing next-generation sequencing that NP binds to select regions throughout the viral genome and is not uniformly bound like ‘beads on a string’ as previously posited. However, it remains unclear how NP, which in vitro has no sequence-specific binding activity towards RNA, localizes to its target sites on vRNA. This specific localization of NP to its target sites on vRNA is essential to maintain viral fitness. Furthermore, a body of evidence suggests that vRNA segments form RNA-RNA interactions between individual segments during the assembly process to facilitate the packaging of the complete genome. Our objectives in this proposal are to examine how NP binding to vRNA changes as the segments progress towards the plasma membrane and how RNA-RNA interactions evolve during transit. The outcome of this proposal will provide a greater understanding of the influenza vRNP structure and the complex mechanism governing influenza vRNP assembly.

影响病毒的组装是一个复杂的协调过程。影响病毒基因组由八个负义的单链RNA片段组成，这些RNA片段被病毒编码的核蛋白（NP）低聚物覆盖。在感染期间，新合成的病毒RNA（VRNA）和NP核心中的核心助理。由VRNA，NP和病毒聚合酶复合酶组成的病毒核糖核蛋白（VRNP）段被转运到质膜，所有八个VRNP段都包装到单个VIRION中。 VRNA和NP之间的相互作用不仅可以保护宿主细胞内的VRNA，而且还促进了VRNP段进入和从细胞核中的运输，VRNA合成以及所有八个VRNA段的选择性包装。我们最近使用需要进行下一代测序的技术显示，NP与整个病毒基因组的选择区域结合，并且不像“弦上的珠子”那样统一地结合，如先前所示。然而，尚不清楚在体外没有序列特异性结合活性与RNA的NP如何定位于vrna上的目标位点。 NP在VRNA上的目标位点的特定定位对于维持病毒适应性至关重要。此外，一大批证据表明，在组装过程中，VRNA片段在各个片段之间形成RNA-RNA相互作用，以促进完整基因组的包装。我们在本提案中的目标是研究NP与VRNA的结合如何随着段向质膜发展而变化以及RNA-RNA相互作用如何在过境过程中发展。该提案的结果将对影响力的VRNP结构和控制影响力VRNP组装的复杂机制提供更多的了解。