Role of Host Angiomotin as a Central Regulator of Filovirus Egress and Dissemination

宿主血管动蛋白作为丝状病毒排出和传播的中央调节剂的作用

基本信息

批准号：
10380684
负责人：
RONALD N HARTY
金额：
$ 22.53万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-03-31 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10380684
关键词：
AMOT gene Animal Model Apoptosis Arenavirus BAG3 gene Binding Biological Biological Assay Biology Cell Culture Techniques Cell physiology Cells Characteristics Complex Disease Disease Outbreaks Dose Ebola Ebola virus Family Filovirus Future Genetic Transcription Goals Host resistance Human In Vitro Infection Lassa virus Length Link Marburgvirus Mediating Mus Nature Oxidoreductase Pathologic Pathway interactions Peritoneal Macrophages Physiological Play Polyubiquitination Proteins RNA Viruses Reporting Role Signal Transduction Systemic infection Tertiary Protein Structure Testing Therapeutic Therapeutic Intervention Tight Junctions Tumor Suppressor Proteins Viral Viral Hemorrhagic Fevers Viral Pathogenesis Virion Virus Virus Diseases Virus Replication Zoonoses cell motility chemokine cytokine emerging pathogen hemorrhagic fever virus in vivo insight member mimicry mouse model new therapeutic target nonhuman primate novel plasma protein Z protein expression protein function ubiquitin-protein ligase

项目摘要

Filoviruses (Ebola [EBOV] and Marburg [MARV]) and arenaviruses (e.g. Lassa virus; LAFV) are zoonotic, emerging pathogens that cause outbreaks of severe hemorrhagic fever in humans and non-human primates. A fundamental understanding of the virus-host interface is critical for developing future strategies and countermeasures for therapeutic intervention. As the filovirus VP40 and LAFV Z matrix proteins drive virion assembly and egress, in part, by hijacking specific host proteins containing WW-domains that can interact with their highly conserved PPxY L-domain motifs, these interactions represent a novel therapeutic target to inhibit egress and dissemination of these hemorrhagic fever viruses. Our early studies to identify host WW-domain proteins that regulate filovirus and arenavirus budding identified the E3 ubiquitin ligases Nedd4, ITCH, and WWP1 as positive regulators of viral PPxY-mediated budding. However, in more recent studies, we identified WW-domain containing proteins YAP/TAZ, BAG3, and now WWOX (WW Domain Containing Oxidoreductase; a multi-functional tumor suppressor), as negative regulators of PPxY-mediated egress of VP40 and Z VLPs. The identification of YAP/TAZ, BAG3, and WWOX as negative regulators of viral PPxY-mediated budding is particularly intriguing since all three of these proteins interact with Angiomotin (Amot), a multi-PPxY containing protein that functions as a “master regulator” of Hippo pathway (YAP/TAZ) signaling, cytoskeletal dynamics, cell migration/proliferation, and tight junction (TJ) integrity. Moreover, expression and stability of Amot itself are regulated by PPxY/WW-domain interactions with the Nedd4 family of E3 ubiquitin ligases. Thus, we hypothesize that Amot is a central interactor linking both the positive and negative WW-domain containing regulators of virus egress, and that modular mimicry between viral and host PPxY motifs and the competitive nature of their binding to the same host WW-domain containing proteins will have a major impact on both cellular processes and viral replication and pathogenesis. Indeed, we were first to report that expression of endogenous Amot positively regulates PPxY-mediated egress of EBOV and MARV VP40 VLPs as well as egress and spread of live EBOV and MARV in cell culture. In this proposal, we will build upon our novel finding that PPxY-containing Amot can positively regulate egress and spread of PPxY-containing viruses including EBOV and MARV, and determine whether the competitive PPxY/WW-domain interplay among VP40/Z – Amot – host WW-domain interactors regulates egress of VLPs and live viruses in vitro and in vivo. Successful completion of these aims will provide novel insights into how this complex network of modular PPxY/WW-domain interactions with Amot impacts late stages of hemorrhagic fever virus egress and dissemination and serve as proof-of principle for therapeutic strategies targeting an essential viral-host interaction that may represent an Achilles heel for numerous RNA viruses.

丝状病毒（埃博拉病毒 [EBOV] 和马尔堡病毒 [MARV]）和沙粒病毒（例如拉沙病毒；LAFV）是人畜共患的新兴病原体，可导致人类和非人类灵长类动物爆发严重出血热。对于未来治疗干预策略和对策的制定至关重要，因为丝状病毒 VP40 和 LAFV Z 基质蛋白在一定程度上驱动病毒粒子组装和排出。通过劫持含有 WW 结构域的特定宿主蛋白，这些蛋白可以与其高度保守的 PPxY L 结构域基序相互作用，这些相互作用代表了抑制这些出血热病毒的流出和传播的新治疗靶点，这是我们鉴定宿主 WW 结构域蛋白的早期研究。调节丝状病毒和沙粒病毒出芽的 E3 泛素连接酶 Nedd4、ITCH 和 WWP1 是病毒 PPxY 介导的正调节因子然而，在最近的研究中，我们发现含有 WW 结构域的蛋白质 YAP/TAZ、BAG3 和现在的 WWOX（含有氧化还原酶的 WW 结构域；一种多功能肿瘤抑制因子）是 PPxY 介导的 VP40 和 VP40 出口的负调节因子。 Z VLPs。YAP/TAZ、BAG3 和 WWOX 作为病毒 PPxY 介导的出芽的负调节因子的鉴定特别令人感兴趣，因为这三种病毒都是如此。蛋白质与血管动蛋白 (Amot) 相互作用，血管动蛋白是一种含有多 PPxY 的蛋白质，可作为 Hippo 通路 (YAP/TAZ) 信号传导、细胞骨架动力学、细胞迁移/增殖以及紧密连接 (TJ) 表达和完整性的“主调节器”。 Amot 本身的稳定性是通过 PPxY/WW 结构域与 E3 泛素连接酶 Nedd4 家族的相互作用来调节的，因此，我们发现 Amot 是一个中心相互作用子。连接包含病毒出口的正向和负向 WW 结构域调节因子，并且病毒和宿主 PPxY 基序之间的模块化模拟以及它们与包含相同宿主 WW 结构域的蛋白质结合的竞争性质将对这两种细胞过程产生重大影响事实上，我们首先报道了内源性 Amot 的表达正向调节 PPxY 介导的 EBOV 和 MARV VP40 VLP 的流出以及活 EBOV 的流出和传播。在本提案中，我们将基于我们的新发现，即含有 PPxY 的 Amot 可以积极调节含有 PPxY 的病毒（包括 EBOV 和 MARV）的流出和传播，并确定 PPxY/WW 结构域之间的竞争性相互作用是否存在。 VP40/Z – Amot – 宿主 WW 域相互作用因子在体外和体内调节 VLP 和活病毒的排出，这些目标的成功完成将为了解如何调节 VLP 和活病毒的流出提供新的见解。这种由模块化 PPxY/WW 结构域与 Amot 相互作用组成的复杂网络会影响出血热病毒的流出和传播的后期阶段，并可作为针对重要的病毒与宿主相互作用的治疗策略的原理证明，这种相互作用可能是许多 RNA 病毒的致命弱点。