Role of host protein GBF1 in organizing enterovirus replication complexes

宿主蛋白 GBF1 在组织肠道病毒复制复合体中的作用

• 批准号: 9295959
• 负责人: George A. Belov
• 金额: $ 37.38万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-06-14 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9295959
• 关键词: Alpha Cell; Antiviral Agents; Ants; Asthma; Biology; Biotin; Brefeldin A; Carrier Proteins; Cell physiology; Cells; Cellular Membrane; Chronic; Clinical; Collaborations; Common Cold; Complex; Coxsackie Viruses; Data; Development; Disease Outbreaks; Encephalitis; Ensure; Enterovirus; Enterovirus 68; Enterovirus 71; FDA approved; Family; Family Picornaviridae; Foundations; Frequencies; GBF1 gene; Genetic Variation; Genome; Golgi Apparatus; Guanine Nucleotide Exchange Factors; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Heart; Human; Human poliovirus; In Vitro; Insulin-Dependent Diabetes Mellitus; Integration Host Factors; Intervention; Knowledge; Label; Life Cycle Stages; Ligase; Lipids; Lung; Macromolecular Complexes; Mediating; Medical; Membrane; Membrane Protein Traffic; Metabolic Pathway; Metabolism; Modeling; Molecular; Molecular and Cellular Biology; Monomeric GTP-Binding Proteins; Mutation; Myocarditis; Myopathy; Neurologic; Obstruction; Organelles; Parasites; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Poliomyelitis; Process; Proteins; Public Health; RNA Viruses; Recruitment Activity; Research; Resistance; Resistance development; Resolution; Rhinovirus; Role; Small RNA; Structure; Syndrome; System; Techniques; Tertiary Protein Structure; Therapeutic; Translations; United States; Vaccination; Vaccines; Vertebrate Viruses; Viral; Viral Proteins; Virus; Virus Assembly; Virus Diseases; Virus Replication; drug development; genetic resource; human disease; in vivo; novel; novel strategies; novel therapeutic intervention; overexpression; pathogen; pressure; protein function; protein transport; scaffold; therapeutic target; trafficking

Description of the project. Viruses are the ultimate intracellular parasites. With minimal genetic resources they are able to reroute cellular metabolic pathways and hijack cellular proteins to promote their own replication. Thus, understanding the role of specific host factors in the viral life cycle is essential for the development of novel anti-viral strategies. Enteroviruses are a group of picornaviruses, small +RNA viruses of vertebrates including humans. Enteroviruses cause clinically and economically important human diseases, ranging from the common cold to fatal encephalitis and myocarditis. High genetic diversity and adaptability of these viruses complicates development of comprehensive vaccines and traditional therapeutics targeting virus-specific proteins. Of all the pathogenic enteroviruses, only polioviruses can be controlled with vaccines, and no clinically approved anti-enterovirus drugs are available. Thus, novel approaches are urgently needed to control enteroviruses associated with human diseases. Picornaviruses replicate their genomes on specialized membrane domains, replication organelles that feature unique lipid and protein composition and whose development relies on the re- organization of cellular lipid synthesis and membrane trafficking pathways. This implies that at least some cellular membrane metabolism components must be indispensable for viral propagation. Indeed, enteroviruses universally require the host protein GBF1 for their replication. GBF1 is a large multi-domain protein that functions as a guanine nucleotide exchange factor (GEF) for select small GTPases of the Arf family. GBF1 is a master coordinator of the early steps of protein transport in the secretory pathway, and participates in maintaining Golgi structure and function and in lipid droplet metabolism. However, how GBF1 supports the viral replication cycle remains unknown. Attempts to relate the known cellular activities of GBF1 such as Arf activation, membrane remodeling and recruitment of cellular proteins to membranes to GBF1 function in viral replication produced controversial results. GBF1 is known to interact with numerous cellular proteins and with the enterovirus replication protein 3A, but how viruses use these interactions to their advantage remains unknown. Herein, we propose a new model in which GBF1 acts as a molecular scaffold to coordinate the assembly of viral and cellular proteins into operational replication complexes. For this project, we built a team of a cell biologist with a superior expertise in GBF1 biology (Sztul), and a virologist with an outstanding background in picornavirus replication (Belov). Together, we will delineate the step(s) in viral life cycle that require GBF1 and uncover the mechanisms of GBF1 action in the formation function of replication complexes. The universal reliance of diverse enteroviruses on GBF1 provides an unprecedented opportunity for the development of broad-spectrum therapeutics targeting GBF1-controlled processes in infected cells.

项目描述。病毒是最终的细胞内寄生虫。具有最小的遗传 他们能够改变细胞代谢途径并劫持细胞蛋白质以促进资源 他们自己的复制。因此，了解特定宿主因素在病毒生命周期中的作用至关重要 用于开发新型抗病毒策略。肠道病毒是一组小核糖核酸病毒，较小 +包括人类在内的脊椎动物的RNA病毒。肠道病毒在临床和经济上引起 重要的人类疾病，从普通感冒到致命的脑炎和心肌炎。高的 这些病毒的遗传多样性和适应性使综合疫苗的开发变得复杂 以及针对病毒特异性蛋白质的传统疗法。在所有致病性肠道病毒中，只有 脊髓灰质炎病毒可以通过疫苗控制，目前尚无临床批准的抗肠道病毒药物 可用的。因此，迫切需要新的方法来控制与人类相关的肠道病毒 疾病。小核糖核酸病毒在专门的膜域上复制其基因组，复制 具有独特脂质和蛋白质组成的细胞器，其发育依赖于重新 细胞脂质合成和膜运输途径的组织。这意味着至少有一些 细胞膜代谢成分对于病毒的繁殖必定是不可缺少的。的确， 肠道病毒普遍需要宿主蛋白 GBF1 来进行复制。 GBF1是一个大型多域 作为 Arf 的特定小 GTP 酶的鸟嘌呤核苷酸交换因子 (GEF) 的蛋白质 家庭。 GBF1 是分泌途径中蛋白质转运早期步骤的主协调员，并且 参与维持高尔基体结构和功能以及脂滴代谢。然而，如何 GBF1 支持病毒复制周期仍然未知。尝试将已知的细胞 GBF1 的活性，例如 Arf 激活、膜重塑和细胞蛋白募集 膜对 GBF1 在病毒复制中的功能的影响产生了有争议的结果。已知 GBF1 会相互作用 具有许多细胞蛋白和肠道病毒复制蛋白 3A，但是病毒如何使用这些蛋白 相互作用对他们有利的情况仍然未知。在此，我们提出了 GBF1 发挥作用的新模型 作为分子支架来协调病毒和细胞蛋白的组装以进行可操作的复制 复合物。对于这个项目，我们建立了一个由在 GBF1 生物学方面拥有卓越专业知识的细胞生物学家组成的团队 （Sztul）和一位在小核糖核酸病毒复制方面具有杰出背景的病毒学家（Belov）。我们一起 将描述病毒生命周期中需要 GBF1 的步骤并揭示 GBF1 的作用机制 复制复合物的形成功能。多种肠道病毒普遍依赖 GBF1为广谱疗法的发展提供了前所未有的机遇 针对受感染细胞中 GBF1 控制的过程。