Defining the contributions of BRCA1, BRCA2, and RAD52 to genome stability

定义 BRCA1、BRCA2 和 RAD52 对基因组稳定性的贡献

基本信息

批准号：
9883062
负责人：
Eric C Greene
金额：
$ 41.09万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-11 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/9883062
关键词：
Address Affect Aging Antineoplastic Agents BARD1 gene BRCA1 Protein BRCA1 gene BRCA2 Mutation BRCA2 gene Binding Biological Assay Cells Characteristics Chromosomal Rearrangement Chromosomes Clinical Complex DNA DNA Double Strand Break DNA Repair DNA Repair Pathway DNA Sequence DNA biosynthesis DNA replication fork Defect Dependence Development Disease Event Exhibits Filament Future Genetic Diseases Genetic Recombination Genome Stability Goals Hereditary Breast and Ovarian Cancer Syndrome Human Human Biology Human Genome Individual Lead Macromolecular Complexes Malignant Neoplasms Mediating Microscopy Molecular Mutate Normal Cell Nucleic Acids Nucleoproteins Optics Outcome Pathway interactions Patients Play Predisposition Process Property Proteins RAD52 gene Reaction Regulation Repair Complex Research Resistance Role Single-Stranded DNA Source Syndrome Time Transact Tumor Suppression Tumor Suppressor Proteins Work anti-cancer therapeutic anticancer research base cancer cell cancer therapy chemotherapy defined contribution experimental study genetic information genome integrity homologous recombination human DNA human disease human imaging inhibitor/antagonist insight malignant breast neoplasm presynaptic prevent protein complex repaired single molecule spatiotemporal therapeutic target time use tool

项目摘要

Project Summary Our chromosomes are continually bombarded with a variety of insults, resulting in damage that must be repaired. By necessity, cells have evolved mechanisms to detect and repair broken strands of DNA, thereby preventing loss of important genetic information. Double-stranded DNA breaks (DSBs) are a type of damage that can result in particularly disastrous outcomes. If not corrected, DSBs can lead to gross chromosomal rearrangements, which are the hallmark of all forms of cancer. Indeed, defects in HR-related proteins are associated with several severe genetic diseases. Patients with these diseases often exhibit a strong predisposition for developing cancers due to a loss of genome integrity. Surprisingly, DNA replication is the primary source of DSBs, and as a consequence rapidly growing cells are especially dependent upon homologous DNA recombination for survival. This dependence upon homologous recombination for the survival of rapidly growing cells highlights the potential for using recombination inhibitors as highly selective anti-cancer therapies. To exploit the clinical potential of homologous recombination inhibitors it will be essential that we more fully understand the molecular underpinnings of the proteins that are involved in regulating and controlling recombination. To help better understand the molecular basis of homologous DNA recombination we have developed powerful new experimental platforms that allow us to directly visualize hundreds of individual DNA molecules at the single molecule level. We are utilizing these unique research tools to probe the fundamental basis for protein-nucleic acid interactions, with emphasis placed upon understanding reactions relevant to human biology and disease. We have used these assays to study human RAD51, which binds to single- stranded DNA forming a key recombination intermediate called the presynaptic complex. Here, we will assess how complexes containing the tumor suppressor protein complexes BRCA1-BARD1, BRCA2- DSS1, and PALB2 promote homologous recombination by regulating the activities of the RAD51 presynaptic complex. We will also examine how the protein RAD52, which newly recognized as an important target for anti-cancer drugs, interacts with RAD51, BRCA1-BARD1, BRCA2-DSS1, and PALB2. We will also study the protein RADX, which is emerging as a key player in genome integrity which functions to downregulate RAD51 activity. We will accomplish these goals by directly visualizing these processes in real-time using optical microscopy. These studies offer the potential for significant new insights into how BRCA1-BARD1, BRCA2-DSS1, PALB2 and RAD52 regulate homologous recombination and support human genome integrity.

项目概要我们的染色体不断受到各种侮辱的轰炸，造成必须予以修复的损害修复了。细胞必然会进化出检测和修复 DNA 断裂链的机制，从而防止重要遗传信息的丢失。双链 DNA 断裂 (DSB) 是一种可能导致特别灾难性后果的损害。如果不加以纠正，DSB 可能会导致严重的后果染色体重排，这是所有形式癌症的标志。事实上，人力资源相关的缺陷蛋白质与几种严重的遗传疾病有关。患有这些疾病的患者通常会表现出由于基因组完整性丧失而极易罹患癌症。令人惊讶的是，DNA 复制是 DSB 的主要来源，因此快速生长的细胞尤其生存依赖于同源DNA重组。这种对同源的依赖重组促进快速生长细胞的存活凸显了利用重组的潜力抑制剂作为高度选择性的抗癌疗法。开发同源的临床潜力重组抑制剂我们必须更全面地了解重组抑制剂的分子基础参与调节和控制重组的蛋白质。为了帮助更好地理解同源 DNA 重组的分子基础，我们开发了强大的新实验平台使我们能够直接可视化数百个个体 DNA 单分子水平上的分子。我们正在利用这些独特的研究工具来探索基本原理蛋白质-核酸相互作用的基础，重点是理解与相关的反应人类生物学和疾病。我们已经使用这些测定法来研究人类 RAD51，它与单链状 DNA 形成关键的重组中间体，称为突触前复合体。在这里，我们将评估含有肿瘤抑制蛋白复合物 BRCA1-BARD1、BRCA2- 的复合物如何 DSS1 和 PALB2 通过调节 RAD51 的活性促进同源重组突触前复合体。我们还将研究蛋白质 RAD52（新近被认为是一种抗癌药物的重要靶点，与 RAD51、BRCA1-BARD1、BRCA2-DSS1 相互作用， PALB2。我们还将研究蛋白质 RADX，它正在成为基因组完整性的关键参与者，具有下调 RAD51 活性的功能。我们将通过直接可视化这些来实现这些目标使用光学显微镜实时处理过程。这些研究为重大新研究提供了潜力深入了解 BRCA1-BARD1、BRCA2-DSS1、PALB2 和 RAD52 如何调节同源重组并支持人类基因组完整性。