Virus-host interactions in frog virus 3 infected cells

青蛙病毒 3 感染细胞中病毒与宿主的相互作用

• 批准号: RGPIN-2015-03989
• 负责人: Brunetti, Craig
• 金额: $ 2.77万
• 依托单位: Trent University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2019
• 资助国家: 加拿大
• 起止时间: 2019-01-01 至 2020-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=687247
• 关键词: Virus; host; interactions; frog; virus

Background: The goal of this proposal is to understand virus-host interactions employed by a relatively unstudied group of large DNA viruses - the Iridoviridae family. Understanding virus-host interacting genes will allow us to explore novel anti-viral strategies, provide new insight into the function of the immune system, and understand how virus infections lead to viral pathogenesis. Although a number of potential Iridoviridae modulators of host anti-viral mechanisms have been identified based on sequence similarity to other viral or cellular genes, few of these genes have been characterized. Therefore, we will use frog virus 3 (FV3), the best-studied member of the Iridoviridae family, to investigate viral modulators of host defense mechanisms. This proposal will characterize the FV3 gene 75L. Previously, our lab showed that 75L binds to LITAF and that the presence of an FV3 infection induces the expression of LITAF.***Hypothesis: We hypothesize that LITAF is part of a conserved cellular defense pathway as its expression is turned on after FV3 infection. We further hypothesize that 75L may antagonize LITAF's function since we have shown that 75L binds to LITAF. In this proposal we will explore whether FV3-75L can inhibit LITAF function. We will also determine whether LITAF represents an uncharacterized cellular host defense mechanism. To achieve these goals, we propose to:***1) Determine if 75L inhibits LITAF function. LITAF has been shown to function in the degradation of a variety of cell surface receptors and induce TNF-a expression. We will determine whether 75L can inhibit these functions.***2) Determine if LITAF affects FV3 replication. Using a variety of assays that monitor different stages of the viral replication cycle, we will determine if viral replication is impaired when 75L protein is reduced or absent.***3) Determine how LITAF is induced upon viral infection. Since LITAF is expressed in response to an FV3 infection, we wanted to determine how an FV3 infection induces LITAF. To test this, we will determine if FV3 induces LITAF through the known pathways that stimulate LITAF expression. ***Significance: The research outlined in this proposal is aimed at characterizing virus-host interacting genes in the Iridoviridae. This proposal focuses on FV3 and its interactions with LITAF. Since the function of LITAF during a viral infection is unknown, our research will not only elucidate the biological function of LITAF, but will also clarify how 75L mediates the inhibition of LITAF. In addition, since mutations in LITAF can cause the human neurodegenerative disease Charcot-Marie Tooth disease (CMT), our continued characterization of LITAF will help in understanding how mutations in LITAF cause CMT and provide insight into potential therapies for this disease.**

背景资料：这项提议的目的是了解一组相对未研究的大型DNA病毒-虹彩病毒科（Iridoviridae）家族所采用的病毒-宿主相互作用。 了解病毒-宿主相互作用基因将使我们能够探索新的抗病毒策略，为免疫系统的功能提供新的见解，并了解病毒感染如何导致病毒发病机制。 尽管已经基于与其他病毒或细胞基因的序列相似性鉴定了宿主抗病毒机制的许多潜在虹彩病毒科调节剂，但是这些基因中很少有被表征。 因此，我们将使用蛙病毒3（FV 3），虹彩病毒科的最好的研究成员，研究宿主防御机制的病毒调节剂。 该提议将表征FV 3基因75 L。 以前，我们的实验室表明75 L与LITAF结合，并且FV 3感染的存在诱导LITAF的表达。假设：我们假设LITAF是保守的细胞防御途径的一部分，因为它的表达在FV 3感染后被打开。我们进一步假设75 L可能拮抗LITAF的功能，因为我们已经表明75 L与LITAF结合。 在本提案中，我们将探索FV 3 - 75 L是否可以抑制LITAF功能。 我们还将确定LITAF是否代表一种未表征的细胞宿主防御机制。 为了实现这些目标，我们建议：*1）确定75 L是否抑制LITAF功能。 已显示LITAF在多种细胞表面受体的降解中起作用并诱导TNF-α表达。 我们将确定75 L是否可以抑制这些功能。2)确定LITAF是否影响FV 3复制。 使用监测病毒复制周期不同阶段的各种测定，我们将确定当75 L蛋白减少或缺失时病毒复制是否受损。3)确定LITAF如何在病毒感染后诱导。 由于LITAF是响应于FV 3感染而表达的，因此我们想确定FV 3感染如何诱导LITAF。 为了测试这一点，我们将确定FV 3是否通过刺激LITAF表达的已知途径诱导LITAF。 * 重要性：本提案中概述的研究旨在表征虹彩病毒科中病毒-宿主相互作用的基因。 该提案侧重于FV 3及其与LITAF的相互作用。 由于LITAF在病毒感染过程中的功能尚不清楚，我们的研究不仅将阐明LITAF的生物学功能，还将阐明75 L如何介导LITAF的抑制作用。 此外，由于LITAF突变可导致人类神经退行性疾病Charcot-Marie Tooth病（CMT），我们对LITAF的持续表征将有助于了解LITAF突变如何导致CMT，并为这种疾病的潜在治疗提供见解。