How does a viral pathogen manipulate DNA Damage Responses to promote infection?

病毒病原体如何操纵 DNA 损伤反应来促进感染？

• 批准号: 10594009
• 负责人: Kinjal Majumder
• 金额: $ 24.88万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-15 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10594009
• 关键词: ATM Signaling Pathway; Animals; Architecture; Area; Binding; Binding Proteins; Binding Sites; CCCTC-binding factor; Cell Nucleus; Cell physiology; Cells; Chromatin; Chromosomes; Complex; DNA; DNA Binding; DNA Damage; DNA Structure; DNA Viruses; DNA damage checkpoint; Disease; Environment; Genome; Genomic Segment; Human; Infection; Knowledge; Malignant Neoplasms; Mediation; Mediator; Mice Minute Virus; NBS1 gene; Nonstructural Protein; Nuclear; Oxidative Stress; Parvovirinae; Parvovirus; Pathogenicity; Phosphorylation; Play; Process; Production; Productivity; Proteins; Reactive Oxygen Species; Research; Role; Signal Pathway; Signal Transduction; Single-Stranded DNA; Site; Stress; System; Training; Transcript; Vertebrate Viruses; Viral; Viral Genome; Virus; Virus Diseases; Virus Replication; Work; biological adaptation to stress; experimental study; gene therapy; insight; mouse genome; novel; pathogen; pathogenic virus; programs; recruit; response; sensor

SUMMARY DNA viruses elicit a cellular DNA damage response (DDR) in infected cells, either in response to incoming viral genomes, or to the large number of foreign genomes generated during virus replication. The DDR forms an innate, critical barrier that can impede or facilitate virus replication. Following host cell infection, replication of the parvovirus minute virus of mice (MVM) induces a sustained DDR, which is then exploited to enhance its replication. Parvovirinae, small non-enveloped icosahedral viruses, are important pathogens in many animal species including humans. These are the only known viruses of vertebrates that contain single-stranded linear DNA genomes, presenting novel replicative DNA structures to the host cell nucleus during infection while relying on cellular processes to replicate. Recent studies by the Pintel lab has shown that MVM interacts with sites of DNA damage to initiate and amplify its infection. This application proposes to examine how MVM exploits the cellular DDR to prepare the nuclear environment for effective parvovirus takeover. Aim 1: How does MVM initiate and activate the host cell’s DNA damage signaling pathway? MVM infection recruits the cellular DDR sensor MRE11 in an MRN (MRE11-RAD50-NBS1) holocomplex- independent manner, but activates a downstream ATM signaling cascade. This project will examine how MRE11 localizes to MVM, and initiates a downstream DDR without its canonical intermediate adaptor complex. Aim 2: How does MVM utilize DNA- and protein- bridging molecules to establish replication centers at sites of DNA damage? MVM associates with cellular DDR sites bound by the architectural protein CTCF, and the DDR-chromatin binding protein MDC1. This study will decipher the mechanism by which CTCF and MDC1 drive the formation of viral replication centers, and how they influence the cellular chromatin architecture. Aim 3: Determine how MVM infection induces additional host-cell DNA damage. MVM infection leads to the production of cellular reactive oxygen species (ROS), which contributes to cellular DNA damage. This research will characterize how MVM induces ROS, and how it utilizes the resulting stress signals to amplify downstream DDR signals, thereby expanding infection. Characterization of the interaction between the incoming viral genome and cellular DDR sensors will provide important insight into how cellular DDR cascades are initiated. Secondly, it will elucidate the mechanisms of trans-interaction between MVM and host chromosome at sites of DNA damage. Finally, studies on ROS induction will help in the understanding of how virus replication induces additional DNA breaks, allowing it to expand further. Examining these processes will be essential for understanding how parvoviruses interact with cells, how they program successful infection and ultimately how they cause disease or persist as gene therapy vehicles. Findings from these studies will be extrapolatable to other DNA viruses and pathogenic viral systems.

概括 DNA病毒引起感染细胞中的细胞DNA损伤反应（DDR），要么是为了响应于 传入的病毒基因组，或在病毒复制过程中产生的大量外国基因组。 DDR形成了一种先天的关键障碍，可以阻碍或促进病毒复制。宿主细胞感染后， 小鼠细小病毒分钟病毒的复制（MVM）诱导了持续的DDR，然后探索至 增强其复制。 Parvovirinae，小的非发育二十体病毒，是许多动物的重要病原体 包括人类在内的物种。这些是唯一含有单链线性的脊椎动物的已知病毒 DNA基因组，在感染期间向宿主细胞核展示新型复制DNA结构 依靠细胞过程来复制。 Pintel Lab的最新研究表明，MVM与 DNA损伤部位启动并扩大其感染。该申请提案要检查MVM 利用细胞DDR来准备核环境，以有效地接管细小病毒。 AIM 1：MVM如何启动和激活宿主细胞的DNA损伤信号通路？ MVM 感染报告了MRN中的细胞DDR传感器MRE11（MRE11-RAD50-NBS1）全胶质体 - 独立的方式，但激活下游的ATM信号级联。该项目将研究如何 MRE11本地化为MVM，并在没有其规范中间适配器复合物的情况下启动下游DDR。 目标2：MVM如何利用DNA和蛋白质桥接分子在建立复制中心 DNA损伤的部位？ MVM与由结构蛋白CTCF约束的细胞DDR位点相关联，并且 DDR-染色质结合蛋白MDC1。这项研究将破译CTCF和MDC1的机制 驱动病毒复制中心的形成，以及它们如何影响细胞染色质结构。 AIM 3：确定MVM感染如何诱导其他宿主细胞DNA损伤。 MVM感染导致 细胞活性氧（ROS）的产生，这有助于细胞DNA损伤。这 研究将表征MVM如何影响ROS，以及它如何利用产生的应力信号放大 下游DDR信号，从而扩大感染。 传入病毒基因组和细胞DDR传感器之间相互作用的表征将提供 关于如何启动细胞DDR级联的重要见解。其次，它将阐明 MVM和宿主染色体之间的跨性别型在DNA损伤部位。最后，研究ROS 诱导将有助于理解病毒复制如何诱导额外的DNA断裂，从而使其能够 进一步扩展。检查这些过程对于理解细小病毒如何与之相互作用至关重要 细胞，如何对成功感染进行编程，以及最终如何引起疾病或持续作为基因治疗 车辆。这些研究的发现将外推到其他DNA病毒和致病性病毒系统。