Role of DNA damage in the early steps of HSV infection and latency in neurons

DNA 损伤在 HSV 感染早期阶段和神经元潜伏期中的作用

• 批准号: 8629026
• 负责人: Matthew D. Weitzman
• 金额: $ 37.3万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-09-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8629026
• 关键词: Ablation; Acyclovir; Address; Afferent Neurons; Animal Model; Antiviral Agents; Behavior; Cell Nucleus; Cells; DNA; DNA Damage; DNA Viruses; DNA repair protein; Data; Dependence; Deposition; Disease; Encephalitis; Environment; Episome; Epithelial Cells; Fingers; Gene Expression; Genes; Genetic; Genome; Goals; Herpesviridae; Herpesvirus 1; Human; Immediate-Early Proteins; Immunohistochemistry; In Situ Hybridization; Infection; Keratitis; Knowledge; Lead; Link; Lytic; Lytic Phase; Maps; Molecular; Mus; Nervous system structure; Neuroglia; Neurons; Nucleosomes; Oligonucleotide Microarrays; Outcome; Peripheral Nervous System; Pharmaceutical Preparations; Positioning Attribute; Presynaptic Terminals; Process; Productivity; Proteins; Recurrence; Recurrent disease; Regulation; Role; Set protein; Signal Transduction; Simplexvirus; Site; Small Interfering RNA; Staging; Stress; Structure of trigeminal ganglion; Surface; Techniques; Therapeutic; Viral; Viral Genome; Viral Proteins; Virus; Virus Diseases; Work; base; cellular targeting; human DNA; in vivo; inhibitor/antagonist; insight; latent infection; lytic replication; mouse model; mutant; novel; pathogen; prevent; protein expression; public health relevance; reactivation from latency; response; stressor; ubiquitin ligase; ubiquitin-protein ligase; ultraviolet damage; virus host interaction

PROJECT SUMMARY Herpes Simplex Virus type 1 (HSV-1) is a human DNA virus that causes a number of diseases involving the human nervous system, including keratitis and encephalitis. The virus is characterized by its ability to form latent infections in neurons of the peripheral nervous system, from which it can reactivate to cause recurrent disease. Our long-term goal is to understand the cellular responses that contribute to establishment of HSV-1 latency specifically in neurons. The latent genome persists in an episomal chromatinized state for the lifetime of the host without detectable protein expression. Reactivation can occur spontaneously or be induced by various forms of stress. The viral immediate early protein ICP0 is a multifunctional protein important for stimulating the initiation of the lytic cycle and also efficient reactivation of latent or quiescent genomes. Exactly how ICP0 creates a favorable cellular environment during lytic replication and promotes reactivation from latency remains unclear, but dependence on the E3 ubiquitin ligase activity of the RING finger domain in ICP0 suggests that degradation of its substrates is involved. Our labs and others have shown that HSV-1 infection activates and exploits aspects of the cellular DNA damage signaling cascade. We have also shown that the DNA damage machinery can act as an intrinsic cellular defense that is inactivated through degradation of specific DNA repair proteins by ICP0. Our central hypothesis is that both the DNA damage response and the behavior of ICP0 are different in neuronal cells, and that these two factors contribute to establishment of latency specifically in neurons. The objective of this proposal is to define the role of the DNA damage response in circularization and nucleosome deposition on the incoming viral genome at the earliest stages of infection, and to determine how this impacts the decision to form lytic or latent HSV infections in neurons. We will examine how ICP0 controls these processes and will determine why the ICP0 protein does not accumulate in the nucleus of infected neurons. The approach will employ novel techniques to determine how cellular responses impact infection in human neurons in culture and in animal models. These neurovirological studies are significant because they are expected to provide insights into virus-host interactions during HSV-1 infection and the early responses that determine the outcome of infection in neurons and non-neuronal cells. Knowledge of HSV-1 neuronal latency will suggest novel targets for developing antivirals against this significant human pathogen.

项目摘要 单纯疱疹1型（HSV-1）是一种人DNA病毒，会引起许多涉及该病毒 人类神经系统，包括角膜炎和脑炎。该病毒的特征是其形成的能力 周围神经系统神经元的潜在感染，它可以从中重新激活以引起复发 疾病。我们的长期目标是了解有助于建立HSV-1的细胞反应 专门在神经元中的延迟。潜在基因组持续存在以偶发性染色体状态 没有可检测到的蛋白质表达的宿主的。重新激活可以自发地发生，也可以通过 各种形式的压力。病毒立即的早期蛋白质ICP0是一种多功能蛋白 刺激裂解周期的启动以及潜在或静止基因组的有效重新激活。确切地 ICP0如何在裂解复制过程中创建有利的细胞环境并促进重新激活 潜伏期尚不清楚，但依赖于ICP0中环指域的E3泛素连接酶活性 表明其底物的降解涉及。我们的实验室和其他实验室表明HSV-1感染 激活并利用细胞DNA损伤信号级联的各个方面。我们还表明了 DNA损伤机械可以充当固有的细胞防御，通过降解而灭活 特定的DNA修复蛋白通过ICP0。我们的中心假设是DNA损伤反应和 ICP0的行为在神经元细胞中是不同的，这两个因素有助于建立 专门在神经元中的延迟。该提议的目的是定义DNA损伤的作用 在最早的最早阶段对传入病毒基因组的循环和核小体沉积的反应 感染，并确定这如何影响在神经元中形成裂解或潜在HSV感染的决定。我们 将检查ICP0如何控制这些过程，并确定为什么ICP0蛋白不累积 在感染神经元的核中。该方法将采用新颖的技术来确定细胞的方式 反应影响文化和动物模型中人类神经元的感染。这些神经病毒学研究 之所以重要，是因为他们有望在HSV-1感染期间对病毒宿主相互作用提供见解 以及决定神经元和非神经元细胞感染结果的早期反应。 HSV-1神经元潜伏期的知识将提出针对此类抗病毒药的新目标 重要的人类病原体。