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The Pox Virion Molecular Interactome

痘病毒体分子相互作用组

基本信息

批准号：
8731174
负责人：
Paul D Gershon
金额：
$ 19.31万
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/8731174
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）：历史上，天花一直是人类的大杀手之一。尽管这种疾病被认为在大约 35 年前就已被根除，但痘病毒仍然是具有医学、生态和农业重要性的一个主要病毒家族。对于人类来说，除了天花重新传入的可能性之外，消灭天花的时间与过去40年非洲和过去10年美国出现人类猴痘的时间同时发生。由于不知道天花的致命因素，此类爆发的全部潜力仍不确定。病毒包膜和衣壳蛋白在介导抗病毒治疗和疫苗作用中的重要性是无可争议的。从小核糖核酸病毒到疱疹病毒，在分子或原子分辨率上对病毒粒子外部分子结构的理解指导了治疗剂的合理设计以及对引起和阻止病毒感染和疾病的机制的理解。然而，由于痘病毒的复杂性、不对称性和异质性，在分子水平上理解其病毒体结构的尝试尤其失败。水平，从而逃避重要的潜在干预途径。 P.I.假设痘苗病毒粒子的复杂性可能被证明是一个致命弱点。此外，对病毒体结构的全面了解可能被视为痘病毒生命周期中最后剩下的黑匣子之一——它影响痘病毒复制的早期转录、基因组脱壳和病毒体组装阶段。我们对痘病毒体结构的了解中的主要漏洞在于病毒体内存在的蛋白质库存之间的水平（这在很大程度上是已知的）以及完整粒子的基本拓扑和形貌特征（也是已知的）。这个介入区域可以被称为病毒粒子的“分子结构”，或蛋白质“相互作用组”。在此 R21 提案中，P.I.选择了与质谱相结合的蛋白质-蛋白质交联方法。这种相互作用组分析方法在许多方面都是公正的，并且在了解复杂细胞组装体的分子结构（例如核孔复合物、20S 蛋白酶体和 RNA 聚合酶 II）方面有着良好的记录。该提案的目标 1 旨在通过共价蛋白质-蛋白质交联/MS，采用“自上而下”（蛋白质水平）和“自下而上”（肽水平）方法来识别病毒粒子核心内直接并置的蛋白质。 P.I.假设痘病毒颗粒核心壁可能与某些有包膜 RNA 病毒的基质蛋白层没有本质上的不同，并且内部深处酶与周围结构蛋白的经典划分可能不像目前假设的那么清晰。