Defining the regulation of double-strand DNA break repair by HIV Vpr

定义 HIV Vpr 对双链 DNA 断裂修复的调节

• 批准号: 10615655
• 负责人: Oliver I Fregoso
• 金额: $ 37.16万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-21 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10615655
• 关键词: Attenuated; BRCA1 gene; Biochemical; Biological Assay; Biology; Cell Cycle; Cell Cycle Arrest; Cells; Chromatin; Clinical; Complex; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Single Strand Break; DNA Viruses; DNA biosynthesis; Data; Dependence; Developing Countries; Development; Drug usage; Ensure; Genes; Genome; Genotoxic Stress; Goals; HIV; HIV-1; Health; Heart; Human; Impairment; In Vitro; Individual; Knowledge; Lentivirus; Life Cycle Stages; Ligation; Malignant Neoplasms; Malignant neoplasm of ovary; Mass Spectrum Analysis; Mediating; Molecular; Molecular Biology; Mutagens; Natural Immunity; Nonhomologous DNA End Joining; Orthologous Gene; Pathogenesis; Pathway interactions; Peripheral Blood Mononuclear Cell; Phenotype; Primate Lentiviruses; Property; Proteins; Proteomics; RNA Viruses; Regulation; Repression; Research; Role; Signal Transduction; Substrate Specificity; Techniques; Testing; Therapeutic; Viral; Viral Proteins; Virus; Virus Replication; Work; antiretroviral therapy; drug repurposing; homologous recombination; human disease; in vivo; malignant breast neoplasm; monocyte; novel; novel therapeutics; protein protein interaction; recombinational repair; recruit; repaired; response; success; therapeutic target; ubiquitin ligase; vpr Genes

PROJECT SUMMARY / ABSTRACT HIV presents a major obstacle to human health, particularly in developing nations. Large strides have been made in understanding the basic molecular biology of HIV, which have led to advances in the development and success of antiretroviral therapies. Yet despite this knowledge, there are still many aspects of HIV biology which we do not understand. If we are to ever truly cure individuals of HIV, we must first fully understand the molecular mechanisms of viral replication to develop novel therapies that take advantage of essential steps in this lifecycle. One such aspect of HIV biology that has remained a mystery despite decades of research is the accessory gene Vpr. Vpr is evolutionarily conserved and important for pathogenesis in vivo, yet no clear role for Vpr in viral replication has been defined. An emerging property of Vpr-associated phenotypes is engagement of the DNA damage response (DDR). The DDR is a signaling cascade that is vital to ensuring the fidelity of the host genome in the presence of genotoxic stress. Growing evidence has emphasized the importance of both activation and repression of the host DDR by diverse DNA and RNA viruses. However, precisely how and why Vpr engages the DDR is unclear. We have recently begun to bridge this gap in knowledge by identifying that Vpr both activates and represses the DDR at multiple steps. Specifically, we have found that Vpr represses the ability of the cell to repair double- strand DNA breaks via homologous recombination (HR) and non-homologous end joining (NHEJ). Based on our preliminary data, we hypothesize that repression of double-strand DNA break repair is central to the primary function of Vpr. Moreover, we propose that the inability of Vpr-expressing cells to repair damaged DNA represents a tractable means to selectively deplete HIV+ cells via synthetic lethality with genotoxic agents that induce low levels of additional DNA damage. We will take a combined molecular, proteomic, and evolutionary approach to directly test our hypotheses. Success of our proposed research will define the primary role of Vpr, it will elucidate how the DDR regulates HIV replication, and it will provide a novel means to treat HIV+ individuals and clear infected cells.

项目总结/摘要 艾滋病毒是人类健康的主要障碍，特别是在发展中国家。我们已经取得了巨大的进步 在了解艾滋病毒的基本分子生物学，这导致了发展和成功的进步， 抗逆转录病毒疗法。然而，尽管有这些知识，艾滋病毒生物学的许多方面仍然是我们所做的。 不明白如果我们要真正治愈艾滋病病毒，我们必须首先充分了解分子生物学。 病毒复制的机制，以开发新的疗法，利用在这个生命周期中的重要步骤。 艾滋病病毒生物学的一个方面，尽管经过几十年的研究，仍然是一个谜，这就是辅助基因 Vpr。Vpr在进化上是保守的，并且对于体内的发病机制是重要的，但是Vpr在病毒感染中没有明确的作用。 已定义复制。Vpr相关表型的一个新特性是DNA的参与， 损伤响应（DDR）。DDR是一种信号级联，对于确保宿主基因组的保真度至关重要 基因毒性应激的情况下。越来越多的证据强调了激活和 通过不同的DNA和RNA病毒抑制宿主DDR。然而，确切地说，Vpr是如何以及为什么 DDR不清楚。 我们最近开始弥合这一知识差距，确定Vpr既激活又抑制细胞内的 多个步骤的DDR。具体来说，我们发现Vpr抑制细胞修复双链的能力。 链DNA通过同源重组（HR）和非同源末端连接（NHEJ）断裂。基于我们 根据初步数据，我们假设双链DNA断裂修复的抑制是原发性 功能Vpr。此外，我们提出，表达Vpr的细胞不能修复受损的DNA， 代表了一种通过遗传毒性剂的合成致死性选择性消耗HIV+细胞的易处理方法， 诱导低水平额外DNA损伤。我们将结合分子、蛋白质组和进化 直接测试我们的假设。我们提出的研究的成功将定义Vpr的主要作用， 它将阐明DDR如何调节HIV复制，并将提供一种治疗HIV阳性个体的新方法 清除感染的细胞