Quantitative mapping of interactions between dengue virus and its hosts

登革热病毒与其宿主之间相互作用的定量图谱

• 批准号: 8918261
• 负责人: Priya Shirish Shah
• 金额: $ 5.8万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8918261
• 关键词: Address; Affinity Chromatography; Arboviruses; Arthropod Vectors; Biochemical Process; Cell physiology; Cells; Characteristics; Chromosome Mapping; Collaborations; Complement; Complex; Coupled; Culicidae; Data; Defect; Dendritic Cells; Dengue; Dengue Infection; Dengue Virus; Distant; Evolution; Genes; Genetic; Genetic study; Habitats; Human; Integration Host Factors; Link; Maps; Mass Spectrum Analysis; Measures; Molecular; Ontology; Pathway interactions; Phenotype; Proteins; RNA Interference; RNA chemical synthesis; Reporter; Reporting; Resistance; Scientist; Staging; System; Therapeutic; Translations; Vertebrates; Virus; Virus Replication; combat; comparative; drug development; functional group; genetic approach; insight; knock-down; monocyte; pathogen; prophylactic; public health relevance; research study; therapeutic development; transmission process; vector mosquito; virus host interaction

DESCRIPTION (provided by applicant): Viruses interact with the host to orchestrate myriad biochemical processes required for replication. A unique characteristic of arboviruses such as dengue virus (DENV), which are transmitted to vertebrates by arthropod vectors, is that these viruses must maintain interactions in two evolutionarily distant hosts to successfully replicate. I this proposal, comparative genetic interaction mapping will be used to identify conserved host factors that are functionally important for DENV replication: Aim 1-Quantitatively map evolutionary constraints of DENV-host interactions A genetic interaction between two factors implies that they are functionally related. Single and pairwise RNAi knockdown coupled with a reporter virus system will be used to quantitatively map genetic interactions of host factors that physically interact with DENV. This will be the first systematic study of genetic interactions in a host-pathogen system. Hierarchical clustering will be applied to the data generated in Aim 1 to identify host factors with similar genetic interaction profiles. Previously acquired DENV-host and host-host protein interaction data will be incorporated to define functional modules. Enrichment analysis will be used to identify functional groups and pathways that are enriched for host replication or restriction factors. The results from Aim 1 will address fundamental questions regarding the evolutionary constraints imposed by hosts on DENV replication. Aim 2-Identify the point of defect in the viral replication cycle for a subset of host replication factors from conserved functional modules Viral replication will be measured at the point of translation, RNA synthesis and exit in both host systems following knockdown by RNAi for 10 host replication factors from conserved functional modules characterized in Aim 1. The experiments in this aim will provide insight into the molecular mechanism of DENV replication in two evolutionarily distant hosts. Ultimately, an evolutionary perspective on DENV-host interactions will be particularly useful for drug development. Targeting conserved or evolutionarily constrained interactions will yield more robust therapies, as DENV will be limited in its ability to evolve resistance.

描述（由申请人提供）：病毒与宿主相互作用，协调复制所需的无数生化过程。登革热病毒（DENV）等虫媒病毒通过节肢动物载体传播给脊椎动物，其独特特征是这些病毒必须在两个进化上相距较远的宿主中维持相互作用才能成功复制。在本提案中，比较遗传相互作用图谱将用于识别对 DENV 复制具有功能重要的保守宿主因子： 目标 1 - 定量绘制 DENV-宿主相互作用的进化限制 两个因子之间的遗传相互作用意味着它们在功能上相关。单一和成对的 RNAi 敲除与报告病毒系统相结合将用于定量绘制与 DENV 物理相互作用的宿主因子的遗传相互作用。这将是第一个针对基因相互作用的系统研究 宿主-病原体系统。层次聚类将应用于目标 1 中生成的数据，以识别具有相似遗传相互作用特征的宿主因子。先前获得的 DENV-宿主和宿主-宿主蛋白质相互作用数据将被纳入定义功能模块。富集分析将用于识别针对宿主复制或限制因子富集的功能组和途径。目标 1 的结果将解决有关宿主对 DENV 复制施加的进化限制的基本问题。目标 2-从保守功能模块中识别宿主复制因子子集的病毒复制周期中的缺陷点 在通过 RNAi 敲除 10 个宿主后，将在两个宿主系统中的翻译、RNA 合成和退出点测量病毒复制来自目标 1 中表征的保守功能模块的复制因子。该目标中的实验将深入了解两个进化距离较远的宿主中 DENV 复制的分子机制。最终，DENV 与宿主相互作用的进化视角对于药物开发特别有用。针对保守或进化受限的相互作用将产生更强大的治疗方法，因为 DENV 进化耐药性的能力将受到限制。