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.

总结 DNA病毒在感染细胞中引起细胞DNA损伤反应（DDR），或者响应于 输入的病毒基因组，或病毒复制过程中产生的大量外源基因组。的 DDR形成一种先天的关键屏障，可以阻止或促进病毒复制。宿主细胞感染后， 小鼠细小病毒微小病毒（MVM）的复制诱导持续的DDR，然后利用DDR 加强其复制。 细小病毒亚科（Parvovirinae）是一类无包膜的二十面体病毒，是许多动物的重要病原 包括人类在内的物种。这些是唯一已知的脊椎动物病毒，含有单链线性 DNA基因组，在感染过程中向宿主细胞核呈现新的复制DNA结构， 依靠细胞过程进行复制Pintel实验室最近的研究表明，MVM与 DNA损伤的位点来启动和扩大其感染。本申请旨在研究MVM如何 利用细胞DDR为有效的细小病毒接管准备核环境。 目的1：MVM如何启动和激活宿主细胞的DNA损伤信号通路？MVM 感染在MRN（MRE 11-RAD 50-NBS 1）全复合物中招募细胞DDR传感器MRE 11， 独立的方式，但激活下游ATM信令级联。本项目将研究如何 MRE 11定位于MVM，并启动下游DDR，而无需其规范的中间衔接子复合物。 目的2：MVM如何利用DNA和蛋白质桥接分子在细胞内建立复制中心？ DNA损伤的部位MVM与由结构蛋白CTCF结合的细胞DDR位点结合， DDR-染色质结合蛋白MDC 1。本研究将阐明CTCF和MDC 1 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。 目的3：确定MVM感染如何诱导额外的宿主细胞DNA损伤。MVM感染导致 细胞活性氧（ROS）的产生，导致细胞DNA损伤。这 研究将描述MVM如何诱导ROS，以及它如何利用由此产生的压力信号放大 下游DDR信号，从而扩大感染。 表征传入病毒基因组和细胞DDR传感器之间的相互作用将提供 重要的洞察细胞DDR级联如何启动。其次，它将阐明 MVM与宿主染色体在DNA损伤位点的反式相互作用。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， 进一步扩大。研究这些过程对于理解细小病毒如何与病毒相互作用至关重要。 细胞，它们如何编程成功感染，以及最终它们如何导致疾病或作为基因治疗持续存在 车辆.这些研究的结果将外推到其他DNA病毒和致病病毒系统。