Essential early events in the flavivirus lifecycle

黄病毒生命周期中重要的早期事件

• 批准号: 10366009
• 负责人: Brett D. Lindenbach
• 金额: $ 41.88万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-09 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366009
• 关键词: ATP phosphohydrolase; Ablation; Address; Antiviral Agents; Antiviral Therapy; Capsid Proteins; Cells; Complement; Culicidae; Data; Dengue; Dengue Virus; Development; Environment; Enzymes; Event; Family; Flavivirus; Future; Gene Expression; Genetic; Genome; Health; Human; Life Cycle Stages; Link; Macromolecular Complexes; Modeling; Molecular; Monitor; Multiprotein Complexes; Nucleocapsid; Peptide Hydrolases; Play; Polyproteins; Process; Protein Family; Proteins; Public Health; RNA replication; Reporter; Role; Serine Protease; Techniques; Translating; Translations; Vaccines; Viral; Viral Genes; Viral Genome; Viral Proteins; Virion; Virus Replication; West Nile virus; Yellow Fever; Yellow fever virus; Zika Virus; arthropod-borne; chemical genetics; combinatorial; design; event cycle; genetic approach; human disease; improved; invertebrate host; member; mutant; prototype; rhomboid; tool; valosin-containing protein; virus host interaction

Project Summary Arthropod-borne flaviviruses such as dengue virus, West Nile virus, yellow fever virus (YFV), and Zika virus, are a major cause of human disease. We are studying the early, post-fusion and pre-replication events in the flavivirus lifecycle by using the prototype flavivirus, YFV, as our model. Our overarching hypothesis is that flaviviruses, which alternately replicate in vertebrate and invertebrate hosts, have evolved to use i) highly conserved factors shared between hosts; and ii) multiple, redundant factors that may not be well conserved between hosts. Therefore, discoveries made with YFV will be validated and studied by comparison to other flaviviruses and between human and mosquito cells. Aim 1 focuses on the post-fusion process of nucleocapsid uncoating, dissecting the mechanisms by which cells unlock this fateful cargo. Aim 1 is a logical extension of our Preliminary Data showing that the delivery of a translatable YFV genome requires cellular ubiquitylation and VCP/p97, a cellular ATPase that extracts ubiquitylated proteins from large macromolecular complexes. Aim 2 focuses on identifying the cellular protease(s) that cleave the viral NS1-2A polyprotein intermediate. Cleavage is essential for flavivirus replication (shown here), yet the identity of this protease has remained elusive for over two decades. We have identified a small family of related candidate NS1-2A proteases and are validating their activity by rigorous, combinatorial genetic ablation. These efforts will solve long- standing puzzles in flavivirus gene expression and replication and identify targets for future development of broadly acting antivirals.

项目概要 节肢动物传播的黄病毒，例如登革热病毒、西尼罗河病毒、黄热病病毒 (YFV) 和寨卡病毒 病毒，是人类疾病的主要原因。我们正在研究早期、融合后和复制前 使用原型黄病毒 YFV 作为我们的模型，黄病毒生命周期中的事件。我们的首要任务 假设黄病毒在脊椎动物和无脊椎动物宿主中交替复制， 进化到使用 i) 宿主之间共享的高度保守的因子； ii) 多重冗余因素 宿主之间可能没有很好地保存。因此，YFV 的发现将 通过与其他黄病毒以及人类和蚊子细胞之间的比较进行验证和研究。 目标 1 重点关注核衣壳脱壳的融合后过程，通过以下方式剖析其机制： 哪些细胞可以解锁这种致命的货物。目标 1 是我们初步数据的逻辑扩展，表明 可翻译 YFV 基因组的传递需要细胞泛素化和 VCP/p97（一种细胞因子） 从大分子复合物中提取泛素化蛋白质的 ATP 酶。目标 2 重点关注 鉴定裂解病毒 NS1-2A 多蛋白中间体的细胞蛋白酶。解理是 对于黄病毒复制至关重要（如图所示），但这种蛋白酶的身份仍然难以捉摸 二十多年来。我们已经确定了一个相关候选 NS1-2A 蛋白酶的小家族，并且 正在通过严格的组合基因消融来验证其活性。这些努力将解决长期 解决黄病毒基因表达和复制中的难题并确定未来的目标 开发广泛作用的抗病毒药物。