Hot Spot Analysis of the Breast Cancer Susceptibility Protein

乳腺癌易感蛋白热点分析

基本信息

批准号：
10356915
负责人：
Deborah F Kelly
金额：
$ 37.01万
依托单位：
PENNSYLVANIA STATE UNIVERSITY, THE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-03-01 至 2025-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10356915
关键词：
3-Dimensional Address Affect Algorithms Architecture Attenuated BRCA1 Mutation BRCA1 Protein BRCA1 gene BRCA2 gene Binding Binding Proteins Binding Sites Biochemical Breast Cancer Cell Breast Cancer cell line Cancer-Predisposing Gene Cell Nucleus Cell Survival Cells Clinical Cryoelectron Microscopy DNA DNA Damage DNA Modification Process DNA Repair DNA Repair Gene DNA lesion Data Deubiquitinating Enzyme Development Disease Disease susceptibility Ensure Estrogens Event Excision Female Genetic Genetic Diseases Genetic Materials Genome Genomic Instability Germ-Line Mutation Hereditary Breast Carcinoma Hereditary Malignant Neoplasm Hot Spot Imaging Device Imaging technology Individual Inherited Knowledge Lead Left Lesion Light Link Malignant Neoplasms Malignant neoplasm of ovary Mass Spectrum Analysis Measurement Measures Metabolic Modification Molecular Molecular Structure Mutate Mutation Nuclear Occupations Oxidative Stress Persons Post-Translational Protein Processing Predisposition Process Property Proteins Publishing Reactive Oxygen Species Research Role Site Structure Tertiary Protein Structure Testing Therapeutic Time Tissues Tumor Suppression Tumor Suppressor Proteins Ubiquitin Woman Work analytical tool biochemical tools brca gene cancer cell carcinogenesis clinically significant confocal imaging early onset experience experimental study genome integrity improved insight lens malignant breast neoplasm mammary operation oxidative damage particle physical property protein structure function public health relevance reconstruction repaired response structural biology targeted treatment three dimensional structure

项目摘要

Abstract Germline mutations in the breast cancer susceptibility gene (BRCA1) are heavily linked to familial breast and ovarian cancers. Women who inherit these mutations are ~60% more likely to develop the disease. During normal female development, critical windows of vulnerability correlate with the early onset of breast cancer. These events coincide with a buildup of DNA lesions in mammary tissue as reactive oxygen species are formed from the metabolic processing of estrogen. Damaged DNA, if left unrepaired, can perpetuate errors in the genome. Reduced expression levels of the BRCA1 protein or BRCA1 mutations, compounded with insufficient lesion repair, provide a tipping point toward cancer induction. At the molecular level, the intricate details of these mechanisms are poorly understood. In the nucleus, BRCA1 associates with its binding partner the BRCA1-Associated Ring Domain protein (BARD1) to help coordinate the repair of DNA modifications. The BRCA1-BARD1 heterodimer performs these operations by interacting with other repair proteins, such as BRCA2, at damaged sites on DNA. In this context, BRCA1 acts as a tumor suppressor to ensure fidelity in the genome. Inherited mutations in BRCA1 can cause functional deficiencies in the protein that affect its job in tumor suppression. Moreover, as decades of scientific research demonstrate BRCA1's multifaceted role in DNA repair, information about the physical properties of BRCA1 are just coming to light. Improving our knowledge of BRCA1's three-dimensional (3D) structure can provide new insights for therapeutic discovery. In the proposed research, we will use a unique combination of cryo-Electron Microscopy (EM) imaging technology and biochemical tools to study the differences in wild type and mutated BRCA1. Our preliminary data suggests that changes in mutated BRCA1 in response to oxidative damage leads to changes in a “modification hot spot” on the protein. We will test this idea across multiple breast cancer cells lines, then develop a new enzymatic approach to attenuate this effect in cancer cells. Our combined biochemical and structural biology strategies will provide a new lens to view the physical nuances of the BRCA1 structure for the first time. This information will shed light on BRCA1 deficiencies relate to cancer susceptibility.

抽象的乳腺癌易感性基因（BRCA1）中的种系突变与家庭乳房和卵巢癌。继承这些突变的妇女患这种疾病的可能性高约60％。期间正常的女性发育，脆弱性的关键窗口与乳腺癌的早期发作相关。这些事件与乳腺组织中的DNA病变相吻合，因为活性氧是由雌激素的代谢加工形成。如果未修复，受损的DNA可能会持续基因组。 BRCA1蛋白或BRCA1突变的表达水平降低，复合病变修复不足，为癌症诱导提供了一个转折点。在分子水平上，复杂的这些机制的细节知之甚少。在细胞核中，BRCA1与其结合伴侣相关与BRCA1相关的环域蛋白（BARD1）有助于协调DNA修饰的修复。这 BRCA1-BARD1异二聚体通过与其他修复蛋白相互作用，例如 BRCA2，在DNA上受损的地点。在这种情况下，BRCA1充当肿瘤抑制剂，以确保在基因组。 BRCA1中的遗传突变会在影响其工作的蛋白质中引起功能缺陷肿瘤抑制。此外，数十年的科学研究表明了BRCA1在 DNA修复，有关BRCA1物理特性的信息刚刚亮起。改善我们的对BRCA1的三维（3D）结构的了解可以为治疗发现提供新的见解。在拟议的研究，我们将使用冷冻电子显微镜（EM）成像的独特组合研究野生型和突变BRCA1差异的技术和生化工具。我们的初步数据表明，突变的BRCA1因氧化损伤而变化导致A的变化蛋白质上的“修饰热点”。我们将在多个乳腺癌细胞线上测试这个想法，然后开发一种新的酶方法来减轻癌细胞中这种影响。我们结合的生化和结构生物学策略将为您提供新的镜头，以查看BRCA1结构的物理细微差别第一次。该信息将阐明与癌症易感性有关的BRCA1缺陷。