Defining the role of Host Hsp70 Subnetworks in Dengue Virus Replication

定义主机 Hsp70 子网在登革热病毒复制中的作用

基本信息

批准号：
9215112
负责人：
JUDITH FRYDMAN
金额：
$ 44.55万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-11-14 至 2021-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9215112
关键词：
ATP Hydrolysis ATP phosphohydrolase Alpha Cell Antiviral Agents Antiviral Therapy Arboviruses Binding Binding Proteins Biochemical Biological Biological Assay Biology Capsid Cells Cellular biology Chemicals Cleaved cell Complex Data Dengue Infection Dengue Virus Dependence Dissociation Enzymes Family Flavivirus Genes Genetic Genome Genomics Goals Guanine Nucleotide Exchange Factors Human Hydrolysis Immunity Immunofluorescence Immunologic Intervention Knowledge Life Cycle Stages Location Mediating Membrane Proteins Molecular Molecular Chaperones Nucleotides Peptide Hydrolases Pharmacology Play Polyproteins Populations at Risk Process Production Protein Isoforms Proteins Proteomics Prothrombin Quality Control RNA RNA Viruses RNA chemical synthesis RNA replication Recruitment Activity Regulation Reporting Resolution Role Specific qualifier value Specificity Structure Substrate Specificity System Tail Tick-Borne Encephalitis Virus Tick-Borne Encephalitis Viruses Toxic effect Ubiquitination Viral Viral Genome Viral Physiology Viral Proteins Virion Virus Virus Diseases Virus Replication West Nile virus Work Yellow fever virus chaperone machinery cofactor experimental study heat-shock proteins 40 inhibitor/antagonist insight knock-down novel particle pathogen prevent protein degradation protein folding protein function proteostasis viral RNA

项目摘要

Defining the role of Host Hsp70 Subnetworks in Dengue Virus Replication Abstract: Viral protein folding and homeostasis depends entirely on the machinery of the host cell. Here we define the dependence of Dengue virus (DENV) on the complex network of Hsp70 chaperones and cochaperones which mediate distinct steps of the virus life cycle. We find that several cytosolic Hsp70 isoforms are required at multiple steps of the viral life cycle to prevent viral protein degradation, promote virion assembly and support viral enzyme function. At each step, Hsp70 function is specified by distinct subnetworks of cofactors, called DnaJs and NEFs, that modulate Hsp70 action and localization at each step. We hypothesize that combinations of DnaJs and NEFs dictate the specific cellular locations, substrate specificities and downstream effectors of Hsp70 in viral replication. We propose to define the mechanism and function of Hsp70 subnetworks in DENV replication through the integration of genetic and proteomic analyses with biochemical and cell biological experiments. Specifically we propose to: (1) Define the mechanism and function of DnaJs in DENV replication; (2) Dissect the role and mechanism of Hsp70 NEFs in DENV replication and (3) Define the mechanism of restriction by the Bag6-centered network. Defining the chaperone subnetworks required for distinct steps of DENV replication will provide new insights into key aspects of the cell biology and molecular mechanism of viral infection. Importantly, Hsp70 provides a susceptible node for antiviral drugs, since compounds inhibiting the Hsp70 cycle blocks DENV infection with negligible toxicity to the host. Our work will thus identify novel targets for pharmacological antiviral intervention and uncover unanticipated cellular mechanisms for viral restriction.

定义宿主HSP70子网在登革热病毒复制中的作用摘要：病毒蛋白折叠和稳态完全取决于宿主细胞的机械。我们在这里定义登革热病毒（DENV）对HSP70伴侣的复杂网络的依赖性和介导病毒生命周期的不同步骤的联合酮。我们发现几种胞质HSP70同工型在病毒生命周期的多个步骤中需要以防止病毒蛋白降解，促进病毒粒子组装并支持病毒酶功能。在每个步骤中，HSP70功能都是由不同的子网指定的称为DNAJS和NEF的辅因子在每个步骤中调节HSP70的作用和定位。我们假设DNAJ和NEF的组合决定了特定的细胞位置，底物 HSP70在病毒复制中的特异性和下游效应子。我们建议定义机制 HSP70子网在DENV复制中通过遗传和蛋白质组学的整合在DENV复制中的功能生化和细胞生物学实验的分析。具体我们建议：（1）定义 DNAJ在DENV复制中的机制和功能；（2）剖析HSP70的作用和机制 DENV复制中的NEF和（3）定义了以BAG6为中心网络的限制机理。定义DENV复制不同步骤所需的伴侣子网将提供新的见解进入病毒感染的细胞生物学和分子机制的关键方面。重要的是，HSP70提供了抗病毒药物的易感淋巴结，因为抑制HSP70周期阻滞DENV感染的化合物与对宿主的毒性微不足道。因此，我们的工作将确定药理学抗病毒干预的新目标并发现意外的病毒限制细胞机制。