Molecular architecture of the Vaccinia virion by structural proteomics

通过结构蛋白质组学研究牛痘病毒粒子的分子结构

基本信息

批准号：
10179428
负责人：
Paul D Gershon
金额：
$ 33.2万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
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：
2019-09-22 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179428
关键词：
3-Dimensional ART protein Africa Agriculture Animals Antiviral Agents Appearance Architecture Biological Assay Capsid Capsid Proteins Cells Chemicals Complex Crosslinker DNA Polymerase II DNA Viruses DNA-Directed RNA Polymerase Data Data Set Disease Disease Outbreaks Docking Drug Targeting Ebola Enzymes Equipment and supply inventories Escherichia coli Exploratory/Developmental Grant for Diagnostic Cancer Imaging Family Family member Future Genes Genome Goals Grant Health Heart Heterogeneity Human Image Immune system In Situ Infection Infusion procedures Intervention Ions Isotope Labeling Knowledge Laboratories Light Linear Programming Mass Spectrum Analysis Mediating Medical Methods Molecular Molecular Structure Monkeypox Morphogenesis Nucleocapsid Pathway interactions Peptides Phenotype Poxviridae Preparation Process Proteins Proteomics Protomer Publishing RNA RNA Viruses Resolution Roentgen Rays Route Small RNA Smallpox Smallpox Vaccine Structural Models Structural Protein Structure Tertiary Protein Structure Testing Therapeutic Time Vaccination Vaccines Vaccinia Vaccinia virus Viral Virion Virus Virus Replication Yeasts ZIKA anthrax lethal factor base bioinformatics tool connectome crosslink design env Gene Products flexibility innovation interest molecular modeling monomer mutant particle protein crosslink protein folding protein structure self organization stoichiometry therapeutic development three dimensional structure tool transmission process virus envelope

项目摘要

The poxviruses comprise a major virus family of medical, ecological and agricultural importance. The most notorious family member, smallpox has been one of the great killers of mankind. Although the disease was eradicated some 40 years ago, the possibility of smallpox re- appearance at some future time has increased immeasurably with the recent demonstration that a poxvirus very similar to smallpox could be recreated de novo, in the laboratory, with ease. Moreover, eradication and the cessation of vaccination has coincided with the appearance of feral human poxviruses including human monkeypox in Africa, the US and UK. Not knowing the lethal factor in smallpox, the full potential of such outbreaks remains uncertain. The importance of virus envelope and capsid proteins in mediating the effects of antiviral therapeutics and vaccines is undisputed. For small RNA viruses in particular, an understanding of virion structure at molecular or atomic resolution has instructed the development of therapeutic agents and an understanding of mechanisms of infection and disease. Due to their complexity, asymmetry and heterogeneity, poxvirus virions have, however, persistently eluded attempts to elucidate their molecular structure, closing a potential avenue of rational design and intervention. The P.I. hypothesizes that the relative complexity of the vaccinia virion may be a therapeutic Achilles heel. Moreover, a molecular-level understanding of virion morphogenesis and organization, one of the last remaining black boxes in the lifecycle of the poxviruses, impinges upon at least five of the seven classical stages of virus replication. A major gap in our knowledge of pox virion structure lies at the level of molecular architecture – an intervening organizational level between ultrastructural features and the inventory of protein molecules contained within the virion. The P.I. has successfully applied a protein-protein chemical crosslinking approach in combination with protein mass spectrometry (XLMS) to discover neighboring proteins and domains within the undisrupted vaccinia virion in situ. Aim 1 of this proposal seeks to deepen the XLMS dataset to a level that will allow protein molecular docking. Combining XLMS with mutant virus particles blocked in morphogenesis and displaying no apparent internal organization, Aim 2 of this proposal asks whether the virion morphogenic pathway follows a classical programmed linear hierarchy or a process of self-organization with no single, dominant route from molecular components to assembled virion. Using “QconCAT” quantitative MS, Aim 3 seeks to convert XLMS data to a molecular model by determining the global stoichiometries of virion proteins.

痘病毒是医学，生态和农业重要性的主要病毒家族。天花最臭名昭著的家庭成员一直是人类的伟大杀手之一。尽管大约40年前，该疾病被放射性化，但天花的可能性可能在最近的某个时间出现，最近的示威表明，可以轻松地在实验室重新创建与天花非常相似的痘病毒。而且，消除和疫苗接种的停止与出现相吻合野性人类痘病毒在内，包括非洲，美国和英国的人类蒙基托克斯。不知道天花中的致命因素，这种暴发的全部潜力仍然不确定。重要性病毒包膜和衣壳蛋白在介导抗病毒疗法和疫苗是无可争议的。特别是对于小的RNA病毒，对病毒结构的理解在分子或原子分辨率上，都指示了治疗剂的发展和了解感染和疾病机制。由于它们的复杂性，不对称性和但是，异质性，痘病毒病毒持续避免了阐明其的尝试分子结构，结束了理性设计和干预的潜在途径。 P.I. 假设离子病毒的相对复杂性可能是治疗性的致命弱点脚跟。此外，对病毒体形态发生和组织的分子水平的理解，一个在痘病毒生命周期中的最后剩下的黑匣子中，至少有五个病毒复制的七个经典阶段。我们对痘病毒知识的主要差距结构在于分子体系结构的水平 - 在组织之间的中间水平超微结构特征和蛋白质分子的清单中包含的病毒素分子。这 P.I.已成功应用了一种蛋白质 - 蛋白质交联方法组合与蛋白质质谱（XLM）一起发现相邻的蛋白质和结构域原位不间断的离甲叶毒素。该提案的目标1旨在将XLMS数据集加深到一个允许蛋白质分支的水平。将XLM与突变病毒颗粒结合在形态发生上阻塞，没有显示明显的内部组织，目标2 提案询问病毒体形态途径是否遵循经典的编程线性层次结构或一个自组织的过程，没有分子的单一占主导地位组装的病毒粒子的组成部分。 AIM 3使用“ QCONCAT”定量MS，旨在转换 XLMS数据通过确定病毒粒子蛋白的全局化学计量来到分子模型。