Structure, dynamics and variation in influenza hemagglutinin-driven fusion

流感血凝素驱动融合的结构、动力学和变化

• 批准号: 9904666
• 负责人: Kelly Keisen Lee
• 金额: $ 39.7万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-04-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904666
• 关键词: Acids; Address; Affect; Animals; Antigenic Variation; Antigens; Antiviral Agents; Architecture; Avian Influenza; Base Sequence; Biological; Biological Process; Biophysical Process; Biophysics; Birds; Cell membrane; Cell-Matrix Junction; Cells; Chimeric Proteins; Collaborations; Cryo-electron tomography; Data; Deuterium; Development; Distal; Docking; Electron Microscopy; Epitopes; Event; Evolution; Exhibits; Foundations; Genetic Materials; Glycoproteins; Goals; H7 hemagglutinin; Hemagglutinin; Human; Hydrogen; Image; Infection; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Influenza Hemagglutinin; Lead; Link; Maps; Mass Spectrum Analysis; Mediating; Membrane; Membrane Fusion; Molecular Conformation; Monitor; Morphology; Mutation; Nature; Pathogenicity; Pathway interactions; Phenotype; Physiological; Point Mutation; Process; Proteins; Reaction; Reassortant Viruses; Research Personnel; Shapes; Site; Specificity; Structure; Surface; Techniques; Testing; Tropism; Vaccines; Variant; Viral; Viral Genome; Virion; Virus; Work; biophysical techniques; capsule; conformational conversion; deep sequencing; design; flu transmission; improved; influenza virus strain; influenza virus vaccine; influenzavirus; inhibitor/antagonist; innovation; insight; mutation screening; particle; pathogenic virus; receptor; receptor binding; transmission process; uptake; virus envelope; virus morphology

Enveloped viruses use specialized protein machinery to fuse their membrane with the membrane of host cells and deliver their genetic material for replication. In influenza virus, the trimeric hemagglutinin (HA) glycoprotein spike is responsible for host cell attachment and membrane fusion. The sequence of events and mechanistic interplay of fusion protein and membranes that lead to fusion have long resisted structural characterization, but through the use of techniques such as cryo-electron tomography (cryo-ET) and structural mass spectrometry that allow native HA and intact virus to be studied under near physiological conditions, we have recently made significant progress in connecting structure, biophysical mechanisms and biological function. Our initial studies focused on the well-characterized X31 H3N2 reassortant virus strain. This work revealed the nature of activation hotspots on the HA fusion protein and elucidated the architecture and sequence of HA-driven membrane remodeling that leads to efficient fusion. The goal of the current proposed project is to apply these powerful approaches to characterize functional variation in HA-mediated membrane fusion by comparing HA from different subtypes and bearing mutations that promote transmission of influenza virus to new hosts. We also will examine the effect of variation in particle morphology on the fusion process, since the M1 matrix protein works together with HA to mediate fusion and it is also the primary determinant of virion morphology. A mechanistic understanding of variations in HA/M1-mediated fusion, linked to a structural framework, will provide a valuable basis for understanding influenza virus pathogenicity, transmission, and evolution. In turn, this information can offer a unique perspective to help guide rational selection of influenza vaccine constructs, immunogen development, and design of HA-targeting inhibitors.

包膜病毒使用专门的蛋白质机制将其膜与宿主细胞膜融合，并传递其遗传物质进行复制。在流感病毒中，三聚体血凝素（HA）糖蛋白刺突负责宿主细胞附着和膜融合。融合蛋白和导致融合的膜的事件序列和机械相互作用长期以来一直抵制结构表征，但通过使用技术，如冷冻电子断层扫描（cryo-ET）和结构质谱，使天然HA和完整的病毒在接近生理条件下进行研究，我们最近在连接结构，生物物理机制和生物功能方面取得了重大进展。我们最初的研究集中在充分表征的X31 H3 N2抗病毒株。这项工作揭示了HA融合蛋白上激活热点的性质，并阐明了导致有效融合的HA驱动的膜重塑的结构和序列。目前提出的项目的目标是应用这些强大的方法来表征HA介导的膜融合中的功能变异，通过比较来自不同亚型的HA和携带促进流感病毒向新宿主传播的突变。我们还将研究颗粒形态变化对融合过程的影响，因为M1基质蛋白与HA一起介导融合，并且它也是病毒体形态的主要决定因素。HA/M1介导的融合的变化，连接到一个结构框架的机制的理解，将提供一个有价值的基础，了解流感病毒的致病性，传播和进化。反过来，这些信息可以提供一个独特的视角，以帮助指导合理选择流感疫苗构建体，免疫原开发和HA靶向抑制剂的设计。

项目成果

Dissection of Epitope-Specific Mechanisms of Neutralization of Influenza Virus by Intact IgG and Fab Fragments.

完整 IgG 和 Fab 片段中和流感病毒的表位特异性机制的剖析。

• DOI: 10.1128/jvi.02006-17
• 发表时间: 2018
• 期刊: Journal of virology
• 影响因子: 5.4
• 作者: Williams,JamesA;Gui,Long;Hom,Nancy;Mileant,Alexander;Lee,KellyK
• 通讯作者: Lee,KellyK

Visualization of conformational changes and membrane remodeling leading to genome delivery by viral class-II fusion machinery.

• DOI: 10.1038/s41467-022-32431-9
• 发表时间: 2022-08-15
• 期刊: Nature communications
• 影响因子: 16.6

Elicitation of Potent Neutralizing Antibody Responses by Designed Protein Nanoparticle Vaccines for SARS-CoV-2.

• DOI: 10.1016/j.cell.2020.10.043
• 发表时间: 2020-11-25
• 期刊: Cell
• 影响因子: 64.5
• 作者: Walls AC;Fiala B;Schäfer A;Wrenn S;Pham MN;Murphy M;Tse LV;Shehata L;O'Connor MA;Chen C;Navarro MJ;Miranda MC;Pettie D;Ravichandran R;Kraft JC;Ogohara C;Palser A;Chalk S;Lee EC;Guerriero K;Kepl E;Chow CM;Sydeman C;Hodge EA;Brown B;Fuller JT;Dinnon KH 3rd;Gralinski LE;Leist SR;Gully KL;Lewis TB;Guttman M;Chu HY;Lee KK;Fuller DH;Baric RS;Kellam P;Carter L;Pepper M;Sheahan TP;Veesler D;King NP
• 通讯作者: King NP

Dissecting Virus Infectious Cycles by Cryo-Electron Microscopy.

• DOI: 10.1371/journal.ppat.1005625
• 发表时间: 2016-06
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Lee KK;Gui L
• 通讯作者: Gui L

How a broadly neutralizing antibody grapples with antigenic and conformational diversity in dengue virus.

广泛中和抗体如何应对登革热病毒的抗原和构象多样性。

• DOI: 10.1016/j.cell.2021.11.020
• 发表时间: 2021
• 期刊: Cell
• 影响因子: 64.5
• 作者: Lee,KellyK