Virus-host interactions in frog virus 3

背景资料：我的研究计划的总体目标是探索青蛙病毒3（FV 3），虹彩病毒科的蛙病毒属的成员中的病毒-宿主相互作用。由于它们对两栖动物群落的影响，Ranaviruses受到关注。两栖动物种群在全球范围内受到威胁，43%的两栖动物表现出种群下降。导致两栖动物数量减少的一个因素是蛙病毒。我们的实验室一直在研究FV 3，以了解该病毒的基本生物学，以及表征FV 3如何影响两栖动物种群。为了了解致病机制和先天免疫防御，我们的实验室将探索病毒与宿主细胞相互作用的两种平行方式。一个研究领域探索病毒-宿主蛋白质-蛋白质相互作用，另一个领域研究基于小信号分子的防御。假设：我们假设FV 3基因97 R和细胞分裂素激素调节宿主细胞中的防御机制。具体而言，我们将鉴定和表征FV 3 97 R的结合伴侣。我们还将确定细胞分裂素在脊椎动物病毒感染中起什么作用。为了实现这些目标，我们建议：1）建立在FV 3感染过程中，prohibitin-1发挥的作用。我们证明了FV 3基因97 R与prohibitin-1结合。我们将确定97 R/prohibitin-1相互作用是否在FV 3感染中发挥生物学作用，可能调节细胞中的细胞死亡途径。2)确定内源性细胞分裂素是否在病毒感染中发生改变。细胞分裂素是在植物中发挥多种作用的激素。我们将探讨FV 3感染是否改变宿主细胞的细胞分裂素的生产，如果是这样，涉及哪些生物学途径。3)确定外源性细胞分裂素是否抑制病毒感染。我们已经证明细胞分裂素可以抑制FV 3感染。我们建议确定FV 3复制周期的哪个阶段被抑制，以及确定抑制FV 3复制的细胞分裂素的完整补体。重要性：该提案中概述的研究旨在利用科学上尚未开发但经济上重要的病毒家族，即虹彩病毒科，探索新的抗病毒策略，为宿主免疫系统的功能提供新的见解，并了解病毒-宿主相互作用如何导致病毒发病机制。本研究重点关注FV 3 97 R及其与prohibitin-1的相互作用，这可能揭示一种新的病毒-宿主相互作用。此外，虽然细胞分裂素激素在植物中有很好的记录，但它们在脊椎动物系统中的作用仍然很大程度上未知。描述脊椎动物中的细胞分裂素将提供对基础生物学的更好理解，并可能揭示未表征的宿主防御机制。最终，这些研究将有助于了解FV 3如何导致疾病，并可能有助于管理自然人群中的病毒爆发。

Background: The overarching goal of my research program is to explore virus-host interactions in frog virus 3 (FV3), a member of the Ranavirus genus of the Iridoviridae family. Ranaviruses are of concern because of their impact on amphibian communities. Amphibian populations are globally threatened with 43% of amphibians demonstrating population declines. One factor contributing to amphibian declines are Ranaviruses. Our lab has been studying FV3 to understand the basic biology of the virus, as well as characterizing how FV3 impacts amphibian populations. To understand mechanisms of pathogenesis and innate immune defenses our lab will explore two parallel ways in which viruses interact with a host cell. One area of investigation explores virus-host protein-protein interactions, the other area studies defences based on small signalling molecules. Hypothesis: We hypothesize that the FV3 gene 97R and cytokinin hormones modulate host defense mechanisms in the cell. Specifically, we will identify and characterize binding partners of FV3 97R. We will also determine what role cytokinins play in vertebrate viral infections. To achieve these goals, we propose to: 1)Establish the role that prohibitin-1 plays during FV3 infections. We demonstrated that the FV3 gene 97R binds to prohibitin-1. We will determine whether the 97R/prohibitin-1 interaction plays a biological role in an FV3 infection, perhaps modulating the cell death pathways in the cell. 2)Ascertain whether endogenous cytokinins are altered in viral infections. Cytokinins are hormones that carry out a variety of roles in plants. We will explore whether an FV3 infection alters the production of cytokinin's by the host cell and if so, what biological pathways are involved. 3)Determine if exogenous cytokinins inhibit viral infections. We have shown that cytokinin's can inhibit an FV3 infection. We propose to determine what stage of the FV3 replication cycle is inhibited as well as determining the full complement of cytokinin's that inhibit FV3 replication. Significance: The research outlined in this proposal is aimed at using a scientifically unexploited but economically important virus family, the Iridoviridae, to explore novel anti-viral strategies, provide new insight into the function of the host immune system, and understand how virus-host interactions lead to viral pathogenesis. This proposal focuses on FV3 97R and its interactions with prohibitin-1, which may shed light on a novel virus-host interaction. In addition, although cytokinin hormones are well documented in plants, their roles in vertebrate systems remain largely unknown. Characterizing cytokinins in vertebrates will provide a better understanding of basic biology and may reveal an uncharacterized host defense mechanism. Ultimately, these studies will help understand how FV3 causes disease and may help management of viral outbreaks in natural populations.