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.

项目的描述。病毒是最终的细胞内寄生虫。遗传最小 他们能够重新布鲁特的细胞代谢途径和劫持蜂窝蛋白来促进 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是必不可少的 为了制定新型的反病毒策略。肠病毒是一组Picornaviruses，小 +包括人类在内的脊椎动物的RNA病毒。肠病毒在临床和经济上引起 重要的人类疾病，范围从普通的冷脑炎和心肌炎。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治疗剂。在所有病原肠病毒中，仅 脊髓灰质炎病毒可以用疫苗控制，并且没有临床批准的抗肠病毒药物是 可用的。因此，迫切需要新的方法来控制与人相关的肠病毒 疾病。 Picornaviruses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传播必须是必不可少的。的确， 肠病毒普遍要求宿主蛋白GBF1复制。 GBF1是一个大型多域 作为选择ARF的小GTPase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运输早期步骤的总协调员， 参与维持高尔基体的结构和功能以及脂质液滴代谢。但是，如何 GBF1支持病毒复制周期仍然未知。尝试关联已知细胞的尝试 GBF1的活性，例如ARF激活，膜重塑和细胞蛋白的募集到 在病毒复制中，gbf1功能的膜产生了有争议的结果。已知GBF1相互作用 具有许多细胞蛋白和肠病毒复制蛋白3a，但是病毒如何使用这些 为了他们的优势，互动仍然未知。在此，我们提出了一个新模型，其中GBF1行动 作为分子支架，以将病毒和细胞蛋白的组装成量复制 复合物。对于这个项目，我们建立了一个细胞生物学家的团队，具有GBF1生物学领域的卓越专业知识 （sztul），以及具有Picornavirus Replication（Belov）背景下杰出背景的病毒学家。在一起，我们 将在需要GBF1的病毒生命周期中描述步骤，并揭示GBF1动作的机制 在复制复合物的形成函数中。各种肠病毒的普遍依赖 GBF1为开发广谱疗法提供了前所未有的机会 靶向受感染细胞中的GBF1控制过程。