Conserved molecular mechanisms of replication for mosquito-borne flaviviruses

蚊媒黄病毒复制的保守分子机制

• 批准号: 10431689
• 负责人: Priya Shirish Shah
• 金额: $ 19.94万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-22 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10431689
• 关键词: Aedes; Affinity Chromatography; Antiviral Therapy; Arboviruses; Arthropod Vectors; Arthropods; Biochemical Process; Bioinformatics; Biophysics; Birds; Cell physiology; Cells; Cessation of life; Characteristics; Culicidae; Data; Dengue Virus; Development; Disease; Distant; Drug Targeting; Engineering; Epidemic; Evolution; Face; Family; Flavivirus; Foundations; Future; Human; Integration Host Factors; Knowledge; Link; Maps; Mass Spectrum Analysis; Measures; Medical; Methods; Molecular; Molecular Target; Pathway interactions; Persons; Pharmacology; Process; Proteins; Proteomics; Public Health; RNA Interference; Refractory; Reproducibility; Risk; Role; Source; Testing; Therapeutic; Vertebrates; Viral; Viral Proteins; Virus; Virus Replication; Work; Yellow fever virus; Zika Virus; biophysical properties; comparative; genetic manipulation; human disease; knock-down; mosquito-borne; novel; obligate intracellular parasite; protein complex; targeted treatment; therapy development; transmission process; vector; vector mosquito

As obligate intracellular parasites, all viruses replicate by coopting host machinery through virus-host protein interactions. Arthropod-borne viruses, which are transmitted to vertebrates by arthropod vectors, must hijack host machinery in human and arthropod cells to accomplish the same fundamental aspects of virus replication. Thus, arthropod-borne viruses maintain protein interactions with host homologs (interologs) to replicate. Identifying these interologs is critical to understanding how an important group of viruses deals with this unique constraint from a biophysical perspective. For flaviviruses transmitted by Aedes mosquitoes (Aedes-borne flaviviruses), it can also inform therapy development by expanding the list of drug targets since these viruses are a major source of human disease. Using a comparative proteomics approach, we recently found large-scale evidence of interologs for dengue virus (DENV), a major Aedes-borne flavivirus that infects nearly 400 million people annually. These interologs involve processes that are essential for virus replication in human and Aedes cells. We hypothesize that Aedes-borne flaviviruses use the conserved interologs to facilitate replication in human and Aedes cells due to the similar constraints place on these viruses and the complexity of maintaining virus-host protein interactions across multiple divergent hosts. The overall objective of this proposal is to systematically compare the role of interologs in virus replication for two Aedes-borne flaviviruses. We will focus on DENV to take advantage of our existing interolog data and yellow fever virus (YFV), a re-emerging Aedes-borne flavivirus that is distantly related to DENV. In Aim 1, we will systematically identify YFV-human and YFV-Aedes protein interactions using affinity purification and mass spectrometry. We will further identify YFV interologs through computational network integration. In Aim 2, we will identify interologs conserved between YFV and DENV using a similar computational network integration approach. We will then test the role of interologs in Aedes-borne flavivirus replication by measuring virus replication following interolog knockdown. This work will identify the conserved molecular mechanisms by which a medically important group of viruses replicates. It will reveal the biophysical parameters that constrain virus evolution. In the future, the interologs we identify could be leveraged as pharmacological or vector engineering targets to inhibit replication of multiple Aedes-borne flaviviruses. Our work would also lay the foundation to search for more interologs conserved across different arthropod vectors and/or different virus families.

作为专性细胞内寄生虫，所有病毒都通过病毒-宿主蛋白相互作用来配合宿主机器进行复制。节肢动物传播的病毒通过节肢动物载体传播到脊椎动物，必须劫持人类和节肢动物细胞中的宿主机器才能完成病毒复制的相同基本方面。因此，节肢动物传播的病毒维持与宿主同源物（间同源物）的蛋白质相互作用以进行复制。识别这些内同源物对于从生物物理学的角度理解一组重要的病毒如何应对这种独特的限制至关重要。对于伊蚊传播的黄病毒（伊蚊传播的黄病毒），它还可以通过扩大药物靶点清单来为治疗开发提供信息，因为这些病毒是人类疾病的主要来源。利用比较蛋白质组学方法，我们最近发现了登革热病毒 (DENV) 内同源物的大规模证据，登革热病毒是一种主要的伊蚊传播黄病毒，每年感染近 4 亿人。这些内源物涉及人类和伊蚊细胞中病毒复制所必需的过程。我们假设伊蚊传播的黄病毒使用保守的内同源物来促进人类和伊蚊细胞中的复制，因为这些病毒受到类似的限制，并且在多个不同宿主之间维持病毒-宿主蛋白相互作用的复杂性。该提案的总体目标是系统地比较内源物在两种伊蚊传播的黄病毒的病毒复制中的作用。我们将重点关注登革热病毒，以利用我们现有的内源数据和黄热病病毒（YFV），黄热病病毒是一种重新出现的伊蚊传播的黄病毒，与登革热病毒有远亲关系。在目标 1 中，我们将使用亲和纯化和质谱系统地鉴定 YFV-人类和 YFV-Aedes 蛋白质相互作用。我们将通过计算网络集成进一步识别 YFV interolog。在目标 2 中，我们将使用类似的计算网络集成方法来识别 YFV 和 DENV 之间保守的 interologs。然后，我们将通过测量 interolog 敲低后的病毒复制来测试 interolog 在伊蚊传播的黄病毒复制中的作用。这项工作将确定一组医学上重要的病毒复制的保守分子机制。它将揭示限制病毒进化的生物物理参数。将来，我们确定的内同源物可以用作药理学或载体工程靶点来抑制多种伊蚊传播的黄病毒的复制。我们的工作还将为寻找更多在不同节肢动物载体和/或不同病毒家族中保守的内同源物奠定基础。