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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：
10414124
负责人：
Nara Lee
金额：
$ 23.82万
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 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10414124
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.

流感病毒的组装是一个复杂的协调过程。流感病毒基因组由八个负义单链RNA片段组成，这些片段被病毒编码的核蛋白(NP)寡聚体包裹。在感染过程中，新合成的病毒RNA(VRNA)与NP在细胞核内结合。病毒核糖核蛋白(VRNP)片段由vRNA、NP和病毒聚合酶复合体组成，被运输到质膜，在质膜上所有8个vRNP片段被包装成单个病毒粒子。VRNA和NP之间的相互作用不仅起到保护宿主细胞内vRNA的作用，而且还促进vRNP片段的进出核、vRNA合成和所有8个vRNA片段的选择性包装。我们最近展示了使用需要下一代测序的技术，NP结合到整个病毒基因组中的选定区域，而不是像之前假设的那样统一结合在一起。然而，NP在体外与RNA没有序列特异性结合活性，它是如何定位到vRNA上的目标位置的，目前尚不清楚。NP在vRNA上的靶点的这种特异性定位对于维持病毒的适合性是必不可少的。此外，大量证据表明，在组装过程中，vRNA片段在单个片段之间形成RNA-RNA相互作用，以促进完整基因组的包装。我们在这个建议中的目标是研究NP与vRNA的结合如何随着片段向质膜的方向发展而变化，以及RNA-RNA相互作用在运输过程中是如何演变的。这项提案的结果将使人们更好地了解流感vRNP的结构和控制流感vRNP组装的复杂机制。