Microscopy & Image Analysis of Unstained Macromolecules

显微镜检查

• 批准号: 7390852
• 负责人: Timothy S Baker
• 金额: $ 44.78万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-06-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7390852
• 关键词: Algae; Animal Viruses; Antibodies; Antibody Binding Sites; Area; Bacteria; Bacteriophages; Biochemical Genetics; Biological Models; Breathing; Capsid; Carbohydrates; Cells; Chlorella; Collaborations; Complex; Cysteine; Cytoplasm; DNA Packaging; Data; Data Collection; Dependence; Dependovirus; Double Stranded DNA Virus; Engineering; Environment; Event; Family; Family Picornaviridae; Fc Receptor; Florida; Fox Chase Cancer Center; Freezing; Frog virus 3; Funding; Geminiviridae; Genetic; Genome; Grant; Head; Hepadnaviridae; Heparin; Homology Modeling; Human; Image; Image Analysis; Imagery; In Situ; In Vitro; Inorganic Sulfates; Insect Viruses; Insecta; Investigation; Iowa; Iridoviridae; Isometric Exercise; Label; Learning; Letters; Life; Life Cycle Stages; Lipid Bilayers; Maleimides; Mammals; Mediating; Membrane; Messenger RNA; Microscopy; Microviridae; Minnesota; Minor; Modeling; Molecular Genetics; Myoviridae; Nebraska; Numbers; Organism; Paramecium; Parasites; Parvoviridae; Penetration; Phycodnaviridae; Platelet-Derived Growth Factor Receptor; Podoviridae; Prolate; Protein Engineering; Proteins; Range; Reoviridae; Reovirus; Research; Resolution; Roentgen Rays; Role; Serotyping; Site; Site-Directed Mutagenesis; Structure; Structure-Activity Relationship; Surface; System; Tail; Techniques; Technology; Testing; Three-Dimensional Image; Three-Dimensional Imaging; Transcript; Transmission Electron Microscopy; Unspecified or Sulfate Ion Sulfates; Viral; Viral Proteins; Virion; Virus; Virus Assembly; Virus Receptors; Work; base; cell attachment protein; desire; improved; insight; macromolecule; man; nanoGold; neutralizing monoclonal antibodies; particle; pathogen; receptor; reconstruction; research study; response

DESCRIPTION (provided by applicant): Viruses are among the best-known and studied pathogens and infect virtually every living organism from bacteria to man. As viruses are parasites of their hosts, the life cycle of any virus is inextricably tied to that of the host cell. Despite this dependence, all viruses share a number of essential tasks, which they must accomplish for survival. A virus must find and recognize a cell in which it can replicate, release its genome into the cell, generate new viral components and assemble these components into precursors that mature into a stable progeny virion which is released from the host cell and transmitted to encounter a new host. Viruses accomplish these tasks in different ways as a result of adaptation to different cellular environments. Each task involves interactions between components within the context of the whole virion and hence requires the visualization of the entire structure at which the techniques of cryo-transmission electron microscopy (cryoTEM) and three-dimensional (3D) image reconstruction ('cryo-reconstruction') excel. We will exploit these techniques to study a diverse range of viruses, including those that infect humans and other mammals, insects, bacteria, and algae. Numerous projects funded by the current grant have illustrated the structural response of different viruses to the common tasks of the viral life cycle. This proposal involves continued as well as new studies that focus on structural investigations of viruses and virus complexes and dynamic events that lie beyond the current realm of crystallographic technology. The large number and extent of our studies are made possible through several fruitful collaborations, which provide important correlative information from biochemical, genetic, and X-ray crystallographic experiments. Our analyses often gain important insights by combining cryo-reconstruction data with available atomic models. Modeling experiments, for example, can provide 'pseudo-atomic' resolution details about the orientation and binding sites of antibody and receptor molecules on viral capsid surfaces that can be tested and refined by means of targeted molecular genetics experiments. Icosahedral as well as non-icosahedral viruses will be studied. These include representatives of several different virus families, all of which are excellent model systems for studying form and function: Myoviridae, Reoviridae, Parvoviridae, Picornaviridae, Podoviridae, Phycodnaviridae, and Iridoviridae.

描述（由申请人提供）：病毒是最著名和研究的病原体之一，几乎感染了从细菌到人的每个生物体。由于病毒是其宿主的寄生虫，因此任何病毒的生命周期都与宿主细胞的生命周期密不可分。尽管有这种依赖性，但所有病毒都共享许多基本任务，它们必须为生存而完成。病毒必须找到并识别一个可以复制的细胞，将其基因组释放到细胞中，产生新的病毒成分并将这些成分组装成前体中，这些成分成熟到稳定的后代病毒体中，从宿主细胞释放并传播到遇到A新主机。 由于适应了不同的细胞环境，病毒以不同的方式完成了这些任务。每个任务都涉及整个病毒率背景下组件之间的相互作用，因此需要可视化整个结构，在该结构中，冷冻传输电子显微镜（crotem）和三维（3D）图像重建（'Cryo-Rectstruction'' ）Excel。我们将利用这些技术来研究各种各样的病毒，包括感染人类和其他哺乳动物，昆虫，细菌和藻类的病毒。当前赠款资助的许多项目都说明了不同病毒对病毒生命周期的常见任务的结构反应。 该建议涉及持续的以及新的研究，这些研究重点是对病毒和病毒复合物的结构研究以及超出当前晶体学技术领域的动态事件。通过几种富有成果的合作，我们的研究的大量和范围成为可能，这些合作提供了来自生化，遗传和X射线晶体学实验的重要相关信息。我们的分析通常通过将冷冻重建数据与可用的原子模型相结合来获得重要的见解。例如，建模实验可以提供有关病毒式衣壳表面上抗体和受体分子的方向和结合位点的“伪原子”分辨率的细节，可以通过有针对性的分子遗传学实验对其进行测试和完善。 将研究二十面体和非染色体病毒。这些包括几个不同病毒家族的代表，所有这些都是研究形式和功能的出色模型系统：Myoviridae，Reoviridae，Parvoviridae，Picornaviridae，Podoviridae，Podoviridae，Phycodnaviridae和Iridoviridae。