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Deciphering the Molecular Assembly Mechanism of Giant DNA Viruses

破译巨型DNA病毒的分子组装机制

基本信息

批准号：
10621855
负责人：
Chuan Xiao
金额：
$ 34.24万
依托单位：
UNIVERSITY OF TEXAS EL PASO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-06-01 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10621855
关键词：
Affect Algorithmic Software Architecture Binding Biological Biological Assay Biophysics Capsid Capsid Proteins Cells Chemistry Cognitive Computer Analysis Cryoelectron Microscopy DNA Viruses Disease Docking Electrons Equation Evolution Fiber Foundations Future Generations Human Image Image Analysis Individual Laboratories Lipids Maps Mathematical Model Simulation Mathematics Methods Minor Modeling Molecular Nobel Prize Pathway interactions Pattern Phase Play Pneumonia Positioning Attribute Process Proteins Publishing Regulation Research Resolution Role Structure Techniques Technology Thick Training Viral Viral Proteins Virion Virus Virus Assembly Virus Diseases Virus-like particle Visualization Visualization software biophysical analysis detector experience experimental study frontier high resolution imaging human disease human pathogen image reconstruction improved insight marine mathematical analysis molecular assembly/self assembly molecular dynamics multimodality nano new therapeutic target novel pathogenic virus prevent protein protein interaction rational design reconstruction self assembly simulation structural biology therapeutic development tool

项目摘要

Project Summary/Abstract Over the last two decades, many giant DNA viruses have been discovered, some of which are bigger than a small cell. How these giant viruses assemble their virion shell from thousands of simple protein building blocks so precisely is a mystery. However, the sheer size of these viruses poses a significant challenge to currently available techniques. This project will tackle this challenge by using a marine giant virus Cafeteria roenbergensis virus (CroV) as a model to decipher the assembly mechanism of giant viruses, and as an opportunity to develop technology to push the resolution limit of these gigantic structures to the atomic level. During the last five years, cryo-electron microscopy (cryo-EM) has become an increasingly powerful tool to study the structures of biological molecules at atomic resolution, earning its developers the 2017 Nobel Prize in Chemistry. We will collect higher quality images using state-of-the-art cryo-EM equipped with latest new hardware, such as energy filters, direct electron detectors and phase plates. Using these images together with new software algorithms, we will determine the structure of giant CroV to high resolution by image analyses and reconstruction. Structures of individual CroV proteins will also be solved to atomic resolution by cryo-EM using various methods and docked into the cryo-EM reconstructed maps. The resultant pseudo-atomic structure will allow characterization of the ultrastructural features and architecture of CroV, building the essential foundation to unravel the assembly of giant viruses. The structure information will be combined with classic biophysical, molecular dynamic simulation, mathematical modeling, and computational analyses to evaluate the novel assembly model of giant viruses. In the new assembly model, the protein shells of giant viruses are assembled continuously from the 5-fold vertices in an interesting spiral way instead of assembled from patches in a step-wise fashion previously assumed. Giant virus protein shell is assembled from protein building block similar to other viruses, including many human pathogens. Some giant viruses have been associated with human diseases such as pneumonia and cognitive functional change. Understanding these principles governing the assembly of giant viruses will improve the development of therapeutic agents to inhibit virus assembly, thus providing a new avenue for preventing and treating viral diseases in general. Elucidation of the molecular interactions that drive assembly of these giant viruses will also shed light on how to control protein-protein interactions effectively, facilitating the rational design of virus-like nanoparticles with a wide size range for biomedical and other nano-applications. Since some giant viruses are bigger than a small cell, techniques and methods developed in this project will push the limits of structural biology and provide new and useful tools to study even larger supramolecular assemblies and eventually the whole cell in the future.

项目总结/摘要在过去的二十年里，人们发现了许多巨大的DNA病毒，其中一些比一个大。小细胞。这些巨大的病毒如何从数千个简单的蛋白质构建模块组装它们的病毒体外壳是个谜然而，这些病毒的庞大规模对目前的研究构成了重大挑战。可用的技术。该项目将通过使用海洋巨型病毒来应对这一挑战。 roenbergensis病毒（CroV）作为一种模型来破译巨型病毒的组装机制，并作为一种有机会开发技术，将这些巨大结构的分辨率限制推到原子水平。在过去的五年中，冷冻电子显微镜（cryo-EM）已成为一种越来越强大的工具，以原子分辨率研究生物分子的结构，为其开发者赢得了2017年诺贝尔奖。化学.我们将收集更高质量的图像使用国家的最先进的冷冻EM配备最新的新硬件，例如能量过滤器、直接电子检测器和相位板。将这些图像与新的软件算法，我们将通过图像分析确定巨大CroV的结构，以获得高分辨率和重建。单个CroV蛋白的结构也将通过cryo-EM解析到原子分辨率使用各种方法并对接到冷冻EM重建图中。生成的伪原子结构将允许表征CroV的超微结构特征和架构，是解开巨型病毒组装的重要基础结构信息将与经典的生物物理、分子动态模拟、数学建模和计算分析，评估巨型病毒的新型组装模型。在新的组装模型中，病毒从5重顶点开始以一种有趣的螺旋方式连续组装，而不是组装从补丁在一个逐步的方式先前假设。巨大的病毒蛋白壳由蛋白质组装而成与其他病毒相似的构建模块，包括许多人类病原体。一些巨型病毒与人类疾病如肺炎和认知功能改变有关。了解这些控制巨型病毒组装的原理将促进治疗剂的发展，病毒组装，从而为预防和治疗病毒性疾病提供了新的途径。阐发驱动这些巨型病毒组装的分子相互作用也将揭示如何控制蛋白质-蛋白质相互作用的有效性，促进了具有宽尺寸的病毒样纳米颗粒的合理设计用于生物医学和其他纳米应用。由于一些巨型病毒比一个小细胞还大，该项目开发的技术和方法将推动结构生物学的极限，并提供新的，这是研究更大的超分子组装体并最终在未来研究整个细胞的有用工具。