Structural Studies of Togaviruses

披膜病毒的结构研究

• 批准号: 10555263
• 负责人: Richard J. Kuhn
• 金额: $ 64.62万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-02-20 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10555263
• 关键词: Africa; Alphavirus; Animal Structures; Antibodies; Antigens; Antiviral Agents; Applications Grants; Architecture; Asia; Binding; Biological; CD209 gene; Capsid Proteins; Cell Surface Receptors; Cells; Chemicals; Chikungunya virus; Clinical; Collaborations; Common Cold Virus; Compensation; Complex; Cryo-electron tomography; Cryoelectron Microscopy; Crystallography; Dedications; Dengue Virus; Detection; Devices; Diamond; Electrons; Engineering; Epidemic; Epitopes; Evolution; Family; Foundations; Funding; Gene Order; Genome; Glycoproteins; Grant; Health; Image; India; Individual; Infection; Investigation; Laboratories; Life Cycle Stages; Lipids; Malaria; Maps; Membrane; Membrane Lipids; Methods; Molecular; Molecular Conformation; Motion; Nature; Paper; Proteins; Publications; RNA; Research; Research Support; Resolution; Rubella; Rubella virus; SLC11A2 gene; Sampling; Site; Southeastern Asia; Stretching; Structure; Surface; Techniques; Technology; Tick-Borne Encephalitis Virus; Togaviridae; Venezuelan Equine Encephalitis Virus; Viral; Virus; Virus-like particle; Work; World Health; Zika Virus; chikungunya; data acquisition; design; detector; env Gene Products; experience; human pathogen; improved; insight; microscopic imaging; neurotropic; neutralizing antibody; new technology; particle; receptor; reconstruction; small molecule; small molecule inhibitor; structural biology; tissue tropism; tomography; tool; vaccine platform

PROJECT SUMMARY Crystallography was the primary structural tool that first opened up structural investigation of viruses at near-atomic resolution. For instance, the first near-atomic resolution structure of an animal virus, namely a common cold virus, was determined by us in the mid-1980s using crystallographic methods (Nature 317:145-153, 1985). However, larger viruses with lipid envelopes are too variable in their structures and have too small a proportion of their surface involved in lattice contacts to make well-ordered crystals. Cryo-electron microscopy has now emerged as the leading tool for advancing structural biology. Single particle averaging of cryo-electron microscopic images is now frequently approaching atomic resolution with the use of direct electron detection devices and technology to compensate for blurring of images due to random motion during data acquisition. However, the limits of this technology are becoming apparent in the investigations of progressively more complex pleomorphic assemblies. Cryo-electron tomography has now emerged as an invaluable tool for the determination of the structure of individual viral particles without reliance on the availability of identical homogeneous particles. However, the resolution of tomographic reconstructions is usually insufficient to determine structures in atomic detail. Thus, a small part of this grant application is dedicated towards extending the resolution of tomographic maps. Alphaviruses and rubella virus constitute the Togavirus family because of their similarity in gene order. Among alphaviruses, Chikungunya virus is a major world health concern because of its re- emergence in Asia and Africa. We plan to extend our previous studies of the icosahedral, lipid- containing alphaviruses, and tackle the more difficult structural analysis of the pleomorphic rubella virus. Specifically, we are planning to obtain the structures of various alphaviruses when complexed with host receptor molecules and with antibodies and small molecule inhibitors that block maturation, attachment and fusion with host cells in order to study the structural intermediates encountered in the viral life cycle. This information will provide viral targets that can be exploited for the design of specific antiviral therapeutics. We are also developing new technology aimed at stretching and pulling individual cryoEM images to make them more similar and homogenous to improve the resolution of tomographic images of pleomorphic biological assemblies such as rubella virus.

项目摘要 晶体学是首先开放的结构研究的主要结构工具 以接近原子分辨率的病毒。例如，动物的第一个近原子分辨率结构 病毒，即一种常见的冷病毒，由我们在1980年代中期使用晶体学确定 方法（自然317：145-153，1985）。但是，具有脂质信封的较大病毒在 它们的结构，其表面的比例太小，参与晶格接触以至于 排序良好的晶体。现在已经出现了冷冻电子显微镜作为前进的领先工具 结构生物学。现在经常 使用直接电子检测设备和技术来接近原子分辨率 补偿数据获取过程中由于随机运动而导致的图像模糊。但是，限制 这项技术在逐渐复杂的调查中变得显而易见 多形组件。 Cryo-Electron层析成像现已成为无价的工具 确定单个病毒颗粒的结构，而不依赖相同的可用性 均匀的颗粒。但是，断层重建的分辨率通常不足 确定原子细节的结构。因此，本赠款申请的一小部分是专门的 旨在扩展层析成像图的分辨率。 α病毒和红宝石病毒构成togavirus家族，因为它们在基因上的相似性 命令。在α病毒中，基孔肯雅病毒是世界卫生的主要关注点，因为它的重新 亚洲和非洲的出现。我们计划扩展我们先前对二十面体脂质的研究 含有α病毒，并解决多形风疹的更困难的结构分析 病毒。具体而言，我们计划在复合时获得各种α病毒的结构 与宿主受体分子以及抗体和小分子抑制剂的结合，阻塞成熟的小分子抑制剂， 与宿主细胞的依恋和融合，以研究在 病毒生命周期。该信息将提供可以利用特定设计的病毒目标 抗病毒疗法。我们还开发了旨在拉伸和拉动的新技术 单个冷冻图像使它们更相似和同质，以提高分辨率 多形生物组件（例如风疹病毒）的层析成像图像。

项目成果

Requirement of a functional ion channel for Sindbis virus glycoprotein transport, CPV-II formation, and efficient virus budding.

• DOI: 10.1371/journal.ppat.1010892
• 发表时间: 2022-10
• 期刊: PLoS pathogens
• 影响因子: 6.7

Assembly, maturation and three-dimensional helical structure of the teratogenic rubella virus.

• DOI: 10.1371/journal.ppat.1006377
• 发表时间: 2017-06
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Mangala Prasad V;Klose T;Rossmann MG
• 通讯作者: Rossmann MG

Development of a Novel Virus-Like Particle Vaccine Platform That Mimics the Immature Form of Alphavirus.

• DOI: 10.1128/cvi.00090-17
• 发表时间: 2017-07
• 期刊: Clinical and vaccine immunology : CVI
• 作者: Urakami A;Sakurai A;Ishikawa M;Yap ML;Flores-Garcia Y;Haseda Y;Aoshi T;Zavala FP;Rossmann MG;Kuno S;Ueno R;Akahata W
• 通讯作者: Akahata W