Structure of a Simple RNA Enveloped Virus

简单 RNA 包膜病毒的结构

• 批准号: 6819806
• 负责人: MICHAEL G ROSSMANN
• 金额: $ 30.47万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6819806
• 关键词: Coronaviridae; Hantavirus; Orthomyxoviridae; RNA virus; Rubivirus; SARS virus; Sindbis virus; X ray crystallography; cryoelectron microscopy; glycoproteins; host organism interaction; interdisciplinary collaboration; protein protein interaction; protein structure function; virus RNA; virus assembly; virus envelope; virus infection mechanism; virus protein

My part of this grant application concerns structural studies of small, single-stranded, RNA viruses and their structural components using cryo=electron microscopy (cryoEM) and X-ray crystallography. The basic concept, which we and others have been exploring in the last decade, is that, whereas larger biological units are generally unlikely to crystallize, their three-dimensional structures can frequently be determined to 9 A resolution or better by cryoEM. At the same time, the atomic structures of some of the component proteins can be determined by X-ray crystallography to near atomic resolution (probably to better than 3 A resolution). These two types of results can be combined to give "quasi-atomic" structures of whole viruses or virus complexes. Although more than half of our effort will be directed to a continuation of our previous investigation of alphaviruses, in particular of Sindbis virus, I plan to extend our efforts to the study of larger and probably less symmetric, enveloped, RNA viruses, including rubella virus (another genus of alphavirus for which it is not clear whether the virions have reproducible icosahedral symmetry), coronaviruses, and possibly also measles virus (an orthomyxovirus) and Hantaan virus (a bunyavirus). The work on alphaviruses will include the determination of the still structurally unknown E2 glycoprotein; extension of the cryoEM map of Sindbis virus to possibly 6 A resolution; study of the immature, pre-maturation, E2-E3 uncleaved virus; analysis of the RNA structure in the core; examinations of the virus bound with cellular receptor molecules and neutralizing antibodies; and further attempts to crystallize Sindbis virus. Work on other ssRNA viruses will need to go hand-in-hand with the development of methods to determine whole virus structure when the virions are asymmetric and pleomorphic, albeit similar in localized surface features. At the same time it will be necessary to start exploring the crystal structures of the component structural proteins. The major emphasis and the major challenge will be the study of the pleomorphic, non-symmetrical enveloped viruses. Tentatively, we plan to use cryoEM tomography to identify regions of local symmetry in the viral membrane and then to average (if appropriate) these regions between different particles. Alternatively, it might be possible to search cryoEM images of a pleomorphic virus for common patterns. These might represent different orientations of a locally recurring structure.

我的赠款应用中的一部分涉及使用冷冻=电子显微镜（冷冻）和X射线晶体学的小型，单链的RNA病毒及其结构成分的结构研究。我们和其他人在过去十年中一直在探索的基本概念是，尽管较大的生物单元通常不太可能结晶，但它们的三维结构通常可以通过冷冻剂确定为9分辨率或更好的分辨率。同时，某些组分蛋白的原子结构可以通过X射线晶体学到接近原子分辨率（可能大于3 A分辨率）来确定。 可以将这两种类型的结果组合起来，从而产生整个病毒或病毒复合物的“准原子”结构。 Although more than half of our effort will be directed to a continuation of our previous investigation of alphaviruses, in particular of Sindbis virus, I plan to extend our efforts to the study of larger and probably less symmetric, enveloped, RNA viruses, including rubella virus (another genus of alphavirus for which it is not clear whether the virions have reproducible icosahedral symmetry), coronaviruses, and possibly also measles virus （一种原病毒）和汉塔病毒（Bunyavirus）。关于α病毒的工作 将包括确定静止的结构未知的E2糖蛋白； sindbis病毒的冷冻图扩展到6个分辨率；研究未成熟，成熟的E2-E3未溶解病毒的研究；分析核心中的RNA结构；与细胞受体分子结合的病毒检查和中和抗体的检查；并进一步尝试结晶信德氏病毒。在其他ssRNA病毒上的工作将需要与方法的开发齐头并进，以确定病毒不对称和多态性的整个病毒结构，尽管在局部表面特征中相似。同时，有必要开始探索组件结构蛋白的晶体结构。 主要的重点和主要挑战将是对多态，​​非及其包膜病毒的研究。暂时，我们计划使用冷冻断层扫描来鉴定病毒膜中局部对称的区域，然后平均（如果适当）不同颗粒之间的这些区域。 另外，可以搜索多形病毒的冷冻图像中的常见模式。这些可能代表局部重复结构的不同方向。