The Pox Virion Molecular Interactome

痘病毒体分子相互作用组

基本信息

批准号：
8582931
负责人：
Paul D Gershon
金额：
$ 21.73万
依托单位：
UNIVERSITY OF CALIFORNIA-IRVINE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-09 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8582931
关键词：
Africa Agriculture Animals Antibodies Antiviral Agents Appearance Applications Grants Architecture Area Binding Boxing Capsid Proteins Complement Complex Core Protein DNA Binding Development Disease Disease Outbreaks Enzymes Equipment and supply inventories Family Family Picornaviridae Future Genetic Transcription Genome Herpesviridae Heterogeneity Human In Situ Intervention Knowledge Lateral Mass Spectrum Analysis Mediating Medical Methodology Molecular Molecular Structure Monkeypox Normal Pressure Hydrocephalus Nuclear Pore Complex Nucleoproteins Nucleosomes Peptides Positioning Attribute Poxviridae Proteins RNA Polymerase II RNA Viruses Resolution Risk Role Route Shapes Smallpox Solubility Staging Structural Protein Structure Techniques Testing Textiles Therapeutic Therapeutic Agents Vaccinia Vaccinium Virion Virus Virus Diseases Yeasts anthrax lethal factor base crosslink design multicatalytic endopeptidase complex novel particle protein complex protein crosslink protein protein interaction public health relevance therapeutic vaccine tool transmission process virus envelope yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Smallpox has, historically, been one of the great killers of mankind. Although this disease is considered to have been eradicated some 35 years ago, the poxviruses, nonetheless, comprise a major family of viruses of medical, ecological and agricultural importance. For humans, aside from the possibility of smallpox re-introduction, eradication has coincided with the appearance of human monkeypox in Africa during the past 40 years and in the US during the past decade. Not knowing the lethal factor in smallpox, the full potential of such outbreaks remains uncertain. The importance of virus envelopes and capsid proteins in mediating the effects of antiviral therapeutics and vaccines is undisputed. From the picornaviruses to the herpesviruses, an understanding of virion outer molecular structures at molecular or atomic resolution, has instructed the rational design of therapeutic agents and an understanding of mechanisms that cause and thwart virus infection and disease. Due to their complexity, asymmetry and heterogeneity, the poxviruses have, however, been particularly persistent in defeating attempts to understand their virion structure at the molecular level, thus evading an important potential avenue for intervention. The P.I. hypothesizes that the complexity of the vaccinia virion may prove to be an Achilles heel. In addition, a full understanding of virion structure may be regarded as one of the last remaining black boxes in the lifecycle of the poxviruses - one which impinges upon the early transcription, genome uncoating and virion assembly stages of poxvirus replication. The major hole in our knowledge of pox virion structure lies at a level between the inventory of proteins present within the virion (which is largely known) and the basic topological and topographical features of the intact particle (also known). This intervening area may be referred to as the virion's "molecular architecture", or protein "interactome". In this R21 proposal, the P.I. has chosen a protein-protein crosslinking approach in combination with mass spectrometry. Such an approach for interactome analysis is unbiased in many respects, and has a track record of informing the molecular architectures of elaborate cellular assemblies such as the nuclear pore complex, 20S proteasome and RNA polymerase II. Aim 1 of this proposal seeks to identify directly juxtaposed proteins within the virion core via covalent protein-protein crosslinking/MS, taking "top-down" (protein-level) and "bottom-up" (peptide-level) approaches. The P.I. hypothesizes that the pox virion core wall may not be fundamentally dissimilar to the matrix protein layers of some enveloped RNA viruses, and that the classical delineation of enzymes in the deep interior with structural proteins surrounding may not be as clear cut as currently supposed.

描述（由申请人提供）：从历史上看，天花一直是人类的伟大杀手之一。尽管大约35年前被认为已经消除了这种疾病，但尽管如此，痘病毒还是构成了医学，生态和农业重要性的主要病毒家族。对于人类而言，除了天花再入学的可能性外，根除与过去40年中非洲人类蒙基托克斯（Monkeypox）的出现相吻合，在过去的十年中。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。毫无疑问，病毒信封和衣壳蛋白在介导抗病毒疗法和疫苗的作用中的重要性。从Picornaviruse到疱疹病毒，对分子或原子分辨率下的病毒体外分子结构的理解都指示了治疗剂的合理设计以及对引起病毒感染和疾病的机制的理解。然而，由于它们的复杂性，不对称和异质性，痘病毒尤其持续到失败的尝试以理解分子的病毒体结构的尝试水平，从而避开了重要的干预潜在途径。 P.I.假设离甲酸的复杂性可能被证明是阿喀琉斯高跟鞋。此外，对病毒体结构的完全理解可能被视为鼠类生命周期中最后一个剩余的黑匣子之一，这种黑匣子会影响早期转录，基因组脱落和病毒式复制阶段。我们对痘病毒结构的知识的主要漏洞在于病毒体中存在的蛋白质库存之间（在很大程度上是已知的）以及完整粒子（也已知）的基本拓扑和地形特征。该中间区域可以称为Virion的“分子结构”或蛋白质“ Interactome”。在此R21提案中已经选择了一种结合质谱法结合使用蛋白质 - 蛋白质交联方法。这种进行相互作用分析的方法在许多方面都公正，并且具有通知详细的细胞组件分子体系结构（例如核孔复合物，20S蛋白酶体和RNA聚合酶II）的记录。该提案的目标1旨在通过共价蛋白质 - 蛋白交联/MS来鉴定病毒核内核直接并置的蛋白质，以“自上而下”（蛋白质级）和“自下而上）和“自下而上”（肽级）方法。 P.I.假设的是，痘病毒核壁可能与某些包裹的RNA病毒的基质蛋白层没有根本不同，并且在深层内部具有结构性蛋白质周围结构蛋白的经典描述可能不会像当前那样清晰。