The RAP80-BRCC36 Deubiquitinating Complex in DNA Repair

DNA 修复中的 RAP80-BRCC36 去泛素化复合物

• 批准号: 10595037
• 负责人: Roger A Greenberg
• 金额: $ 32.18万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-12-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10595037
• 关键词: ATM Gene Mutation; ATM deficient; Affect; Affinity; BARD1 gene; BRCA mutations; BRCA1 gene; Binding; Binding Proteins; Biochemical; Cancer Etiology; Cells; Chromatin; Clinical; Collaborations; Complex; DNA; DNA Damage; DNA Double Strand Break; DNA Repair; DNA Replication Damage; DNA replication fork; Deubiquitinating Enzyme; Elements; Etiology; Event; Excision; Functional disorder; Generations; Genetic; Genetically Engineered Mouse; Germ-Line Mutation; Goals; Hematopoietic stem cells; Hereditary Breast and Ovarian Cancer Syndrome; Histones; Hydrolysis; Hypersensitivity; Investigation; Knockout Mice; Link; Lysine; Malignant Neoplasms; Malignant neoplasm of ovary; Mass Spectrum Analysis; Mediating; Methodology; Modality; Modification; Molecular; Monitor; Mutation; Nonhomologous DNA End Joining; Nucleosomes; Null Lymphocytes; Other Genetics; Outcome; Pennsylvania; Poly(ADP-ribose) Polymerase Inhibitor; Poly(ADP-ribose) Polymerases; Post-Translational Protein Processing; Predisposition; Proteins; Proteome; Proteomics; Recombinants; Research; Resistance; Role; Single-Stranded DNA; Site; Structure; Surface; System; Testing; Therapeutic; Topoisomerase-I Inhibitor; Ubiquitin; Western Blotting; Work; ataxia telangiectasia mutated protein; candidate identification; cell type; dimer; enzyme activity; gene network; genome integrity; histone modification; homologous recombination; in vivo; insight; loss of function mutation; malignant breast neoplasm; member; novel; prevent; programs; reconstitution; response; stem cell function; structural biology; targeted agent; treatment response; ubiquitin isopeptidase

Contact PD/PI: Greenberg, Roger A. Summary Faithful execution of homologous recombination DNA repair is essential for genome integrity and is a critical determinant of cancer etiology and therapeutic response. This relationship is prominently exemplified by hereditary breast and ovarian cancer, which arises as a consequence of germline mutations to BRCA1 and a network of genes encoding its interacting partners. Dysfunction within this BRCA network produces hypersensitivity to poly(ADP)ribose polymerase inhibitors, which are clinically approved agents that have efficacy in the setting of BRCA mutation and homologous recombination deficiency in general. Resistance frequently occurs to these agents as a consequence of restored homologous recombination DNA repair. It is thus essential to understand how this occurs at a fundamental mechanistic level. We discovered that BRCA1 is targeted to lysine63-linked ubiquitin chromatin regions aligning DNA double-strand breaks and damaged replication forks. This localization requires BRCA1 interaction with the A-complex. This dimeric complex of five proteins includes a ubiquitin binding protein, RAP80, and a deubiquitinating enzyme, BRCC36. RAP80 and BRCC36 are specific for binding lysine63-linked ubiquitin and its hydrolysis. Deficiency of the A-complex reduces BRCA1 DNA damage localization and causes excessive end resection at replication fork damage. Interestingly, loss of the A-complex confers resistance to PARP inhibitors in genetic backgrounds where end resection is diminished. How the BRCA1-A-complex links chromatin recognition to control of end resection and therapeutic response is not currently understood. This proposal will integrate in vivo, cellular, and structure guided biochemical approaches to understand how the A-complex links DNA double-strand break ubiquitin recognition on nucleosomes to homologous recombination. To achieve these goals, we develop novel methodologies that enable us to identify the entire DNA damage response proteome on chromatin and how this affects nucleosome modifications and stability. These studies will yield fundamental advances to the understanding of genome integrity control and clarify new strategies to target underlying vulnerabilities in a broad range of malignancies. ! !

联系PD/PI：Greenberg，RogerA。 概括 同源重组DNA修复的忠实执行对于基因组完整性至关重要，这是关键 癌症病因和治疗反应的决定因素。这种关系由 遗传性乳腺癌和卵巢癌，是由于BRCA1的种系突变而引起的 编码其相互作用伙伴的基因网络。该BRCA网络中的功能障碍产生 对聚（ADP）核糖聚合酶抑制剂的高敏性，这些抑制剂是具有疗效的临床认可的剂 在BRCA突变和同源重组缺乏症的情况下。电阻经常 由于恢复的同源重组DNA修复而发生，这些药物发生。因此，这是必不可少的 了解这是如何在基本机械级别上发生的。 我们发现BRCA1针对赖氨酸63连接的泛素染色质区域对齐DNA双链 断裂和损坏的复制叉。该定位需要BRCA1与A复合物的相互作用。这 五种蛋白的二聚体复合物包括泛素结合蛋白Rap80和去泛素化酶， BRCC36。 RAP80和BRCC36特定于结合赖氨酸63连接的泛素及其水解。不足 A复合物减少了BRCA1 DNA损伤的定位，并在复制叉时导致过度终端切除 损害。有趣的是，在遗传背景下，A复合物的丧失赋予对PARP抑制剂的抗性 终端切除术减少。 BRCA1-A复合物如何将染色质识别链接到控制终端切除 目前尚不了解治疗反应。 该建议将在体内，细胞和结构中整合引导的生化方法，以了解如何了解 A-复合物将核小体对泛素识别的DNA双链断裂与同源重组。 为了实现这些目标，我们开发了新的方法，使我们能够识别整个DNA损害 染色质的响应蛋白质组及其如何影响核小体的修饰和稳定性。这些研究会 从理解基因组完整性控制并阐明针对​​目标的新策略方面的基本进步 在广泛的恶性肿瘤中的根本漏洞。 呢 呢