Novel regulations of DNA damage repair

DNA损伤修复的新调控

基本信息

批准号：
9326453
负责人：
Junjie Chen
金额：
$ 36.6万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
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/9326453
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 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 Outcome Pathway interactions Patients Play Poly(ADP-ribose) Polymerases Process Proteins Radiation Recruitment Activity 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 killings malignant breast neoplasm mutant neoplastic cell novel p53-binding protein 1 recombinational repair repaired resistance mechanism response senescence targeted cancer therapy therapy resistant treatment response tumorigenesis

项目摘要

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.

项目概要 DNA 损伤反应和 DNA 修复的缺陷是基因组不稳定和肿瘤发生。更好地了解 DNA 修复所涉及的途径不仅可以提高我们的研究能力对癌症病因的了解，也为癌症治疗提供了新的靶点。参与的关键蛋白质 DNA修复和肿瘤发生是p53结合蛋白1，即53BP1。我的实验室多年来一直致力于 53BP1。我们小组是最先展示的小组之一 53BP1 在 DNA 损伤反应中的作用。我们建立了第一批 53bp1 基因敲除 (KO) 小鼠；我们也揭示 53BP1 是 DNA 修复所必需的，并在体内充当肿瘤抑制因子。此外，我们阐明了 DNA 损伤后 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 53BP1 在 DNA 断裂位点的募集和积累。然而，53BP1 也可以以不依赖于 H2AX 的方式定位到 DNA 损伤位点，尽管根本机制仍有待确定。此外，我们表明 53BP1 对于特定的修复过程称为类别转换重组，表明 53BP1 参与了特殊的 DNA 修复与典型的非同源末端连接途径明显不同的途径。我们最近的研究和其他人的研究表明，53BP1 抑制 BRCA1 缺陷的同源重组修复细胞，这对于对基于聚（ADP-核糖）聚合酶抑制剂的癌症的反应至关重要疗法。总之，这些数据强调了 53BP1 在对抗同源重组中的重要性针对 DNA 损伤进行修复。在本提案中，我们计划重点关注 53BP1 并从分子水平上进行阐明。水平在 DNA 损伤后如何调节 53BP1 并有助于 DNA 修复和基因组维护。为了进一步了解 53BP1 在 DNA 损伤位点的定位和功能的调控，我们最近进行串联亲和纯化并结合质谱分析来鉴定能够与 53BP1 的区域特异性相关，该区域对于其定位到 DNA 是必要且充分的损坏地点。令人惊讶的是，我们发现了几种新型 53BP1 结合蛋白，包括 NUDT16、 NUDT16L1 和 DEK。在本提案中，我们将1）进一步确定NUDT16L1和NUDT16在 53BP1 调节和 DNA 损伤反应，2) 阐明 DEK 和其他新发现的 53BP1 相关蛋白在损伤诱导的 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发挥作用的关键成分 53BP1 的上游并与 53BP1 一起在 DNA 修复和基因组维护中发挥作用。