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: 10563191
• 负责人: Pradeep D Uchil
• 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/10563191
• 关键词: ATP phosphohydrolase; Ablation; Address; Antiviral Agents; Antiviral Therapy; Capsid Proteins; Cells; Complement; Culicidae; Data; 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 virus; Zika Virus; arthropod-borne; chemical genetics; combinatorial; design; 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.

项目摘要 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Zika 病毒是人类疾病的主要原因。我们正在研究早期，融合后和复习 通过使用原型Flavivirus YFV作为我们的模型，黄素生命周期中的事件。我们的总体 假设是，在脊椎动物和无脊椎动物宿主中交替复制的黄素病毒具有 演变为使用i）主机之间共享的高度保守因素； ii）多个冗余因素 在主机之间可能无法很好地保守。因此，用YFV做出的发现将是 通过与其他黄素和人类和蚊子之间的比较进行了验证和研究。 AIM 1专注于核素脱落后的融合后过程，剖析机制 哪些细胞解锁了这一命运的货物。 AIM 1是我们的初步数据的逻辑扩展 可翻译的YFV基因组的递送需要细胞泛素化和VCP/p97（细胞） 从大型大分子复合物中提取泛素化蛋白的ATPase。 AIM 2专注于 识别裂解病毒NS1-2A多蛋白中间体的细胞蛋白酶。乳沟是 对于黄病毒的复制至关重要（此处显示），但是这种蛋白酶的身份仍然难以捉摸 超过二十年。我们已经确定了一小部分相关候选人NS1-2A蛋白酶和 通过严格的组合遗传消融来验证其活性。这些努力将解决长期 在黄病毒基因表达和复制中站立难题，并确定未来的目标 开发广泛作用的抗病毒药。