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.

丝病毒（埃博拉病毒[EBOV]和MARBUG [MARV]）是人畜共患病，新兴的病原体会导致零星和 人类和非人类灵长类动物的严重出血热暴发。就像Ebov和Marv可以 在中枢神经系统，精液，眼睛和其他免疫学特权地点中建立持续感染 进行抗体疗法及其重新出现会导致长期后遗症和死亡，对宿主的理解 可能导致新抗病毒疗法发展的防御机制比以往任何时候都更为重要。我们的 实验室是识别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基质蛋白是病毒组装和出口的驱动力，用于调节病毒的晚期阶段 复制。为了更好地理解功能性的filevirus-Host Interactome，我们的实验室专注于直接 PPXY晚期（L）域基序之间的相互作用在EBOV的VP40蛋白（EVP40）和 MARV（MVP40）和来自选定宿主蛋白的模块化WW域。除了filevirus的积极调节剂 出口，我们最近确定了对VP40介导的萌芽的负调节的WW-domain相互作用者， 包括YAP/TAZ，BAG3和WWOX。尽管这些负调节器的影响可能是多因素的，并且 复杂的是，我们进行了一个有趣的观察，即Filamin B是YAP/TAZ的常见下游效应子 和Bag3，是实时EBOV/MARV感染/进入和Live Ebov/Marv Egress的潜在双调节器。 由于犯罪病毒生命周期相对端的关键阶段的双重角色调节是新颖且出乎意料的，因此 该提案的最终目标是确定宿主蛋白丝蛋白B如何机械影响丝状病毒 生命周期。 Filamins是肌动蛋白交联/稳定蛋白的家族，可调节血浆肌动蛋白动力学 控制细胞迁移和形态，囊泡运输，大型细胞增多症和细胞分裂的膜。 值得注意的是，大型细胞增多症 大刺体 是 （MPS）与肌动蛋白，丝蛋白和丝蛋白相关的蛋白密度密集 当这些病毒进入囊泡中时，丝状病毒的关键进入途径 称为 。此外， 丝蛋白与模仿病毒体形成和/或出口（例如丝状形成）的细胞过程有关， 细胞迁移和细胞因子。因此，我们假设Filamin B调节了丝状病毒的感染性/进入 通过对大型细胞增多症的影响，并通过对丝状病毒样的影响来调节丝状病毒。 丝状病毒颗粒。在此探索性建议中，我们建议研究丝胺的机制 调节丝状病毒感染/进入（AIM 1）和出口（AIM 2）。这些研究的结果将通过 识别一种新型宿主蛋白，该蛋白的表达调节了丝状病毒中的两个关键和相反的步骤 生命周期；进入和出口，并提供对功能性货币托管宿主的更好的基本了解 相互作用和宿主防御机制，可能导致新的抗病毒疗法的发展。