Novel regulations of DNA damage repair

DNA损伤修复的新调控

• 批准号: 9883637
• 负责人: Junjie Chen
• 金额: $ 43.64万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-16 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9883637
• 关键词: Affinity Chromatography; BRCA1 gene; Binding Proteins; Cancer Etiology; Cancer Patient; Cell Death; Cell Nucleus; Cells; Chromatin; Clinical; Coupled; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Repair Gene; DNA Repair Pathway; DNA damage checkpoint; DNA lesion; Data; Defect; Double Strand Break Repair; Genes; Genome; Genomic Instability; Goals; Human; Immunoglobulin Class Switching; Immunoglobulin Switch Recombination; Knockout Mice; Laboratories; Lead; Maintenance; Malignant Neoplasms; Malignant neoplasm of ovary; Mass Spectrum Analysis; Mediating; Molecular; Mutagenesis; Mutation; Nonhomologous DNA End Joining; Outcome; Pathway interactions; Play; Poly(ADP-ribose) Polymerases; Process; Proteins; Radiation; Regulation; Resistance; Role; Signal Pathway; Site; Source; Sun Exposure; Tumor Suppressor Proteins; base; cancer therapy; chemotherapeutic agent; driving force; genome integrity; homologous recombination; in vivo; inhibitor/antagonist; malignant breast neoplasm; mutant; neoplastic cell; novel; p53-binding protein 1; patient response; recombinational repair; recruit; repaired; resistance mechanism; response; senescence; targeted cancer therapy; therapy resistant; treatment response; tumorigenesis

PROJECT SUMMARY Defects in DNA damage response and DNA repair are the driving forces of genomic instability and tumorigenesis. Gaining a better understanding of the pathways involved in DNA repair not only increases our understanding of cancer etiology, but also provides new targets for cancer therapies. A key protein involved in DNA repair and tumorigenesis is p53-binding protein 1, i.e. 53BP1. My laboratory has been working on 53BP1 for many years. Our group was one of the first to demonstrate the role of 53BP1 in DNA damage response. We established the first 53bp1 knockout (KO) mice; we also revealed that 53BP1 is required for DNA repair and acts as a tumor suppressor in vivo. In addition, we elucidated the regulation of 53BP1 after DNA damage. We and others demonstrated that the H2AX-dependent DNA damage signaling pathway controls the recruitment and accumulation of 53BP1 at sites of DNA breaks. However, 53BP1 can also localize to DNA damage sites in an H2AX-independent manner, although the underlying mechanisms remain to be determined. Moreover, we showed that 53BP1 is critical for a particular repair process called class-switch recombination, indicating that 53BP1 is involved in a special DNA repair pathway that is distinctly different from the canonical nonhomologous end-joining pathway. Our recent studies and those of others suggest that 53BP1 suppresses homologous recombination repair in BRCA1-deficient cells, which is critically important for response to poly (ADP-ribose) polymerase inhibitor-based cancer therapies. Together, these data highlight the importance of 53BP1 in counteracting homologous recombination repair in response to DNA damage. In this proposal, we plan to focus on 53BP1 and elucidate at the molecular level how 53BP1 is regulated after DNA damage and contributes to DNA repair and genome maintenance. To further understand the regulation of 53BP1 localization and function at DNA damage sites, we recently performed tandem affinity purification coupled with mass spectrometry analysis to identify proteins that would specifically associate with a region of 53BP1, which is necessary and sufficient for its localization to DNA damage sites. Surprisingly, we uncovered several novel 53BP1-binding proteins, including NUDT16, NUDT16L1, and DEK. In this proposal, we will 1) further determine the roles of NUDT16L1 and NUDT16 in 53BP1 regulation and in the DNA damage response, and 2) elucidate the functional significance of DEK and other newly discovered 53BP1-associated proteins in damage-induced 53BP1 localization, DNA repair, and the maintenance of genomic integrity. These studies will help us understand the key components that act upstream of 53BP1 and function together with 53BP1 in DNA repair and genome maintenance.

项目摘要 DNA损伤反应和DNA修复中的缺陷是基因组不稳定性的驱动力和 肿瘤发生。更好地了解DNA修复中涉及的途径不仅增加了我们的 了解癌症病因，但也为癌症疗法提供了新的靶标。涉及的关键蛋白 DNA修复和肿瘤发生是p53结合蛋白1，即53BP1。 我的实验室已经在53BP1上工作了很多年。我们的小组是第一个展示的人之一 53BP1在DNA损伤反应中的作用。我们建立了第一只53BP1敲除（KO）小鼠；我们也是 表明DNA修复需要53BP1，并且在体内充当肿瘤抑制器。另外，我们 DNA损伤后阐明了53BP1的调节。我们和其他人证明了H2AX依赖性 DNA损伤信号通路控制DNA断裂部位的53BP1的募集和积累。 但是，53BP1也可以以H2AX独立的方式定位于DNA损伤位点，尽管 潜在机制仍有待确定。此外，我们表明53BP1对于特定 维修过程称为类开关重组，表明53BP1参与了特殊的DNA修复 与规范的非同源最终连接途径有明显不同的途径。我们最近的研究 其他的表明53BP1抑制了BRCA1缺陷的同源重组修复 细胞，这对于对聚（ADP-核糖）聚合酶抑制剂癌症的反应至关重要 疗法。这些数据共同强调了53BP1在抵消同源重组中的重要性 响应DNA损伤而修复。在此提案中，我们计划专注于53BP1并在分子上阐明 在DNA损伤后如何调节53BP1的水平，并有助于DNA修复和基因组维持。 为了进一步了解DNA损伤位点的53BP1定位和功能的调节，我们最近 进行了串联亲和力纯化，加上质谱分析，以鉴定蛋白质 特别与53BP1区域相关，这是必要且足以将其定位到DNA 损坏站点。令人惊讶的是，我们发现了包括Nudt16在内的几种新颖的53BP1结合蛋白 NUDT16L1和DEK。在此提案中，我们将进一步确定Nudt16l1和Nudt16在 53BP1调节和DNA损伤反应，2）阐明了DEK和 其他新发现的53BP1相关蛋白在损伤引起的53BP1定位，DNA修复和 维持基因组完整性。这些研究将帮助我们了解行动的关键组成部分 在DNA修复和基因组维持中，在53BP1上游并与53BP1一起功能。