Dissecting the Functional Domains of Infected Cell Protein 0 of Herpes Simplex Virus 1

剖析单纯疱疹病毒 1 感染细胞蛋白 0 的功能域

• 批准号: 9238640
• 负责人: Haidong Gu
• 金额: $ 37.72万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-15 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9238640
• 关键词: Acute Promyelocytic Leukemia; Adhesions; Adult; Age; Applications Grants; Bacterial Artificial Chromosomes; Binding; Biology; Cell Line; Cells; Clinical; Communicable Diseases; Complex; Consensus; Consensus Sequence; Corneal Ulcer; DNA Viruses; Data; Defense Mechanisms; Development; Disease; Elements; Ensure; Equilibrium; Ganglia; Gene Expression; Genes; Genetic study; Genital system; Genomics; Goals; Growth; Health; Herpes Labialis; Herpes encephalitis; Herpesvirus 1; Host Defense; Host Defense Mechanism; Human; Immediate-Early Proteins; Immunity; Immunocompetent; Immunocompromised Host; Individual; Infection; Knowledge; Laboratories; Life Cycle Stages; Lytic Phase; Measures; Mediating; Mission; Molecular; Newborn Infant; Nuclear; Orthologous Gene; Outcome; Pathogenesis; Pathogenicity; Pathway interactions; Play; Population; Process; Protein Isoforms; Proteins; Publications; Publishing; Recurrence; Regulation; Reporting; Repression; Role; Simplexvirus; Structure; System; Testing; Viral; Viral Genes; Virus; Virus Diseases; War; base; gene product; gene repression; genetic regulatory protein; in vivo; latent infection; mutant; neurotropic virus; novel therapeutic intervention; novel therapeutics; pathogen; prevent; prophylactic; protein purification; public health relevance; reactivation from latency; ubiquitin-protein ligase; viral DNA; virus host interaction

 DESCRIPTION (provided by applicant): Herpes simplex virus 1 (HSV-1) is a neurotropic virus that infects over 70% of the world adult population. It establishes a lifelong latency in trigemina ganglia and reactivates sporadically to spread to naïve population at their young age. It causes a wide range of clinical manifestations such as cold sores, genital ulceration, keratitis, and the deadly herpes encephalitis. Infected cell protein no. 0 (ICP0), an immediate early protein that is a key regulator for both lytic infection and latency reactivation, plays a vital role in the counteractions against host anti-viral defenses. ICP0 contains an E3 ubiquitin ligase, which mediates proteasomal degradation of several host restrictive factors and therefore enhance viral gene expression. It also interacts with numerous cellular proteins in order to regulate a diverse array of cell pathways participating in anti-viral defenses. The multiple functions of ICP0 located in different domains of ICP0 must coordinate with each other to orchestrate a robust infection. Results from our lab and several other labs have shown that a dynamic nuclear domain 10 (ND10) structure undertakes a close interplay with ICP0 in the tug-of-war of HSV-1 infection. On one hand, ND10 converges at viral DNA for gene repression. On the other hand, ICP0 merges with ND10 and degrades its organizer protein PML (promyelocytic leukemia) in order to disperse ND10 bodies and release the repression. We have previously reported that ICP0-ND10 interaction is a dynamic process that includes sequential steps of adhesion, fusion and retention, and specific ICP0 domains are involved in the faithful execution of these dynamic steps. The overall objective of the present application is to dissect the functional domains of ICP0 that are important for ND10 fusion process and E3 ubiquitin ligase activity, and to delineate ICP0 domain coordination in its counteractions against the host defense. We plan to construct ICP0 mutant viruses to identify functional domains, to characterize the coordination among different domains, and to isolate proteins interacting to these specific domains. We intend to pursue the following specific aims in this grant proposal: (i) to delineate the molecular basis of ND10 fusion process and (ii) to characterize the regulatory mechanisms of ICP0 E3 ubiquitin ligase. These studies are highly relevant to the mission of understanding infectious diseases caused by HSV-1. The outcomes of these proposed studies will greatly advance our knowledge of the initiation of HSV-1 infection, as well as the molecular mechanisms of host defenses. This study is expected to have a significant impact in developing new anti-herpes therapies and in understanding cellular defense mechanisms against DNA viruses.

 描述（由申请人提供）：单纯疱疹病毒1型（HSV-1）是一种嗜神经病毒，感染世界70%以上的成年人群。它在三叉神经节中建立了终身潜伏期，并偶尔重新激活，在年轻时传播给幼稚人群。它引起广泛的临床表现，如唇疱疹、生殖器溃疡、角膜炎和致命的疱疹性脑炎。感染细胞蛋白0号（ICP 0）是一种立即早期蛋白，是裂解性感染和潜伏期再激活的关键调节因子，在对抗宿主抗病毒防御中起着至关重要的作用。ICP 0含有E3泛素连接酶，其介导几种宿主限制性因子的蛋白酶体降解，从而增强病毒基因表达。它还与许多细胞蛋白相互作用，以调节参与抗病毒防御的多种细胞途径。ICP 0的多种功能位于 在ICP 0的不同域中，必须相互协调以协调强有力的感染。我们实验室和其他几个实验室的结果表明，动态核结构域10（ND 10）结构在HSV-1感染的拔河中与ICP 0密切相互作用。一方面，ND 10在病毒DNA处收敛，进行基因阻遏。另一方面，ICP 0与ND 10融合并降解其组织者蛋白PML（promyelocytic leukemia，早幼粒细胞白血病），以分散ND 10小体并释放抑制。我们以前曾报道，ICP 0-ND 10相互作用是一个动态的过程，包括粘附，融合和保留的顺序步骤，和特定的ICP 0域参与忠实地执行这些动态步骤。本申请的总体目标是剖析对ND 10融合过程和E3泛素连接酶活性重要的ICP 0的功能结构域，并描述ICP 0结构域在其对抗宿主防御的对抗中的协调。我们计划构建ICP 0突变体病毒，以确定功能结构域，表征不同结构域之间的协调，并分离与这些特定结构域相互作用的蛋白质。我们打算在这项拨款申请中实现以下具体目标：（i）描绘分子 ND 10融合过程的基础和（ii）表征ICP 0 E3泛素连接酶的调控机制。这些研究与理解HSV-1引起的传染病的使命高度相关。这些研究的结果将极大地推进我们对HSV-1感染的启动以及宿主防御的分子机制的认识。这项研究预计将对开发新的抗疱疹疗法和了解针对DNA病毒的细胞防御机制产生重大影响。