Role of Host Filamin Proteins in Regulating Filovirus Entry and Egress

宿主细丝蛋白在调节丝状病毒进入和排出中的作用

• 批准号: 10644499
• 负责人: RONALD N HARTY
• 金额: $ 26.83万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-10 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10644499
• 关键词: Actins; Affect; Antibody Therapy; Antiviral Therapy; BAG3 gene; Biochemical; Biological Assay; Cell Fractionation; Cell Separation; Cell division; Cell membrane; Cell physiology; Cells; Cellular Morphology; Cessation of life; Complex; Cytokinesis; Development; Disease Outbreaks; Ebola; Ebola virus; Eye; Family; Filopodia; Filovirus; Future; Goals; HIV-1; Host Defense Mechanism; Human; Immunologics; Infection; Integrins; Laboratories; Life Cycle Stages; Link; Liquid substance; Magnetism; Mammalian Cell; Marburgvirus; Mediating; Membrane; Microscopy; Molecular; Pathway interactions; Process; Proteins; Proteome; Reagent; Recombinants; Regulation; Ribonucleases; Role; Seminal fluid; Site; Small Interfering RNA; Testing; Therapeutic Intervention; Total Internal Reflection Fluorescent; Vesicle; Vesicular stomatitis Indiana virus; Viral; Viral Hemorrhagic Fevers; Virion; Virus; Virus Assembly; Virus Diseases; Virus Replication; Zoonoses; cell motility; chronic infection; combat; confocal imaging; crosslink; driving force; emerging pathogen; extracellular; filamin; insight; link protein; long-term sequelae; member; nonhuman primate; novel; particle; protein crosslink; trafficking; ubiquitin-protein ligase; virus host interaction

Filoviruses (Ebola [EBOV] and Marburg [MARV]) are zoonotic, emerging pathogens that cause sporadic and global outbreaks of severe hemorrhagic fever in humans and non-human primates. As EBOV and MARV can establish persistent infections in the CNS, semen, eye, and other immunologically privileged sites inaccessible to antibody therapy, and their re-emergence can result in long-term sequelae and death, understanding of host defense mechanisms that could lead to the development of new antiviral therapies is more critical than ever. Our lab is at the forefront of identifying host proteins that interact both physically and functionally with the filovirus VP40 matrix protein, the driving force for virus assembly and egress, to regulate these late stages of virus replication. To better understand the functional filovirus-host interactome, our laboratory has focused on direct interactions between the PPxY Late (L) domain motif conserved in the VP40 proteins of EBOV (eVP40) and MARV (mVP40) and modular WW-domains from select host proteins. In addition to positive regulators of filovirus egress, we have recently identified WW-domain interactors that negatively regulate VP40-mediated budding, including YAP/TAZ, BAG3, and WWOX. While the impact of these negative regulators is likely multifactorial and complex, we have made the intriguing observation that filamin B, a common downstream effector of YAP/TAZ and BAG3, acts as a potential dual regulator of live EBOV/MARV infectivity/entry and live EBOV/MARV egress. As dual role regulation of key stages at opposite ends of the filovirus lifecycle is novel and unexpected, the ultimate goal of this proposal is to determine how host protein filamin B mechanistically impacts the filovirus lifecycle. Filamins are a family of actin crosslinking/stabilizing proteins that regulate actin dynamics at the plasma membrane to control cell migration and morphology, vesicle trafficking, macropinocytosis, and cell division. Notably, macropinocytosis macropinosomes is (MPs) that are densely coated with actin, filamin, and filamin-associated proteins a key entry pathway for filoviruses, as these viruses enter the cell in vesicles termed . In addition, filamins are linked to cellular processes that mimic virion formation and/or egress such as filopodia formation, cell migration, and cytokinesis. Thus, we hypothesize that filamin B regulates infectivity/entry of filoviruses through effects on macropinocytosis, and regulates filovirus budding through effects on egress of filopodia-like filovirus particles. In this exploratory proposal, we propose to investigate the mechanisms by which filamin regulates filovirus infectivity/entry (Aim 1) and egress (Aim 2). Results from these studies will impact the field by identifying a novel host protein(s) whose expression modulates two critical and opposite steps in the filovirus lifecycle; entry and exit, as well as providing a better fundamental understanding of the functional filovirus-host interactome and host defense mechanisms that could lead to the development of new antiviral therapies.

丝状病毒（埃博拉病毒 [EBOV] 和马尔堡病毒 [MARV]）是人畜共患的新兴病原体，可导致散发性和 全球爆发人类和非人类灵长类动物严重出血热。由于 EBOV 和 MARV 可以 在中枢神经系统、精液、眼睛和其他难以进入的免疫特权部位建立持续感染 抗体疗法，它们的重新出现可能导致长期后遗症和死亡，了解宿主 可能导致新抗病毒疗法开发的防御机制比以往任何时候都更加重要。我们的 实验室处于鉴定与丝状病毒在物理和功能上相互作用的宿主蛋白的最前沿 VP40 基质蛋白，病毒组装和排出的驱动力，调节病毒的这些后期阶段 复制。为了更好地了解功能性丝状病毒-宿主相互作用组，我们的实验室重点关注直接 EBOV (eVP40) VP40 蛋白中保守的 PPxY Late (L) 结构域基序与 MARV (mVP40) 和来自选定宿主蛋白的模块化 WW 结构域。除了丝状病毒的正调节因子 出口，我们最近发现了 WW 域相互作用子，它可以负向调节 VP40 介导的出芽， 包括 YAP/TAZ、BAG3 和 WWOX。虽然这些负面监管机构的影响可能是多因素的， 复杂的情况下，我们进行了有趣的观察，Filamin B 是 YAP/TAZ 的常见下游效应子 和 BAG3，充当活 EBOV/MARV 感染性/进入和活 EBOV/MARV 出口的潜在双重调节器。 由于丝状病毒生命周期两端关键阶段的双重作用调节是新颖且出乎意料的， 该提案的最终目标是确定宿主蛋白细丝蛋白 B 如何机械地影响丝状病毒 生命周期。细丝蛋白是肌动蛋白交联/稳定蛋白家族，可调节血浆肌动蛋白动力学 膜控制细胞迁移和形态、囊泡运输、巨胞饮作用和细胞分裂。 值得注意的是，巨胞饮作用 大胞质体 是 (MP) 紧密包裹有肌动蛋白、细丝蛋白和细丝蛋白相关蛋白 丝状病毒的关键进入途径，因为这些病毒通过囊泡进入细胞 称为 。此外， 细丝蛋白与模拟病毒颗粒形成和/或排出的细胞过程有关，例如丝状伪足形成， 细胞迁移和胞质分裂。因此，我们假设细丝蛋白 B 调节丝状病毒的感染性/进入 通过影响巨胞饮作用，并通过影响丝状伪足样的出口来调节丝状病毒出芽 丝状病毒颗粒。在这个探索性提案中，我们建议研究细丝蛋白的机制 调节丝状病毒感染性/进入（目标 1）和外出（目标 2）。这些研究的结果将影响该领域 鉴定一种新的宿主蛋白，其表达调节丝状病毒中两个关键且相反的步骤 生命周期；进入和退出，以及提供对功能性丝状病毒宿主的更好的基本了解 相互作用组和宿主防御机制可能导致新的抗病毒疗法的开发。