Phospho-Binding Ligands and Substrates of BRCA1

BRCA1 的磷酸结合配体和底物

基本信息

批准号：
8502497
负责人：
MICHAEL B YAFFE
金额：
$ 19.66万
依托单位：
MASSACHUSETTS INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8502497
关键词：
BARD1 gene BRCA1 Mutation BRCA1 gene Binding Binding Proteins Biological Markers Breast C-terminal Cell Cycle Progression Cells Collaborations Complementary DNA Complex DNA Damage DNA Double Strand Break DNA Repair Environment Estrogens Exposure to Genotoxic Stress Goals Incidence Laboratories Ligand Binding Malignant Neoplasms Mammary Gland Parenchyma Mammary gland Mediating Methods Mitotic spindle Molecular Mutation Nature Organ Outcome Ovarian Ovary Phosphoserine Play Post-Translational Protein Processing Protein Binding Proteins Proteome Proteomics Research Risk Role Screening Result Signal Pathway Signal Transduction Susceptibility Gene Threonine Tissues Tumor Suppression Tumor Suppressor Proteins Ubiquitination Validation Woman Work Xenobiotics base cancer risk carcinogenesis drug development environmental agent experience genome wide association study genome-wide improved insight malignant breast neoplasm new technology novel prevent protein protein interaction response screening tumor ubiquitin ligase ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): In an ever more industrialized environment, we are increasingly exposed to a wide range of xenobiotic compounds, many of which are DNA damaging agents that have contributed to an increasing incidence of cancer. Improving our understanding of how cells respond to and repair DNA damage will allow us to better assess genotoxic stress upon exposure to environmental agents and to develop novel molecular strategies to improve cellular responses to DNA damage. In the present proposal we focus on the mechanism responsible for breast and ovarian-specific tumor suppression by the Breast Cancer-1 susceptibility gene product (BRCA1), a key molecule involved in DNA damage signaling. We previously identified the BRCT domains of BRCA1 as phosphoserine/threonine-binding domains, and this phospho-binding function has recently been shown to be critical for BRCA1's tumor suppressor function. We will conduct a comprehensive proteome-wide screen to identify direct BRCT domain-dependent binding partners of BRCA1 and downstream targets for the ubiquitin ligase activity of BRCA1 in complex with its binding partner BARD1 in estrogen-responsive tissues. The proposed studies fit well within the long term goal of our laboratory to understand, in molecular detail, how post-translational modifications by cell signaling pathways, together with modular binding domains, regulate multiple aspects of the cellular response to DNA damage and cell cycle progression. The results of the screen should provide (1) a comprehensive unbiased identification of all BRCA1-interacting proteins in estrogen-responsive breast cells, along with determination of whether they are or are not substrates for BRCA1-BARD1 ubiquitin ligase activity; (2) the identification of potential targets and biomarkers of increased risk for DNA damage-induced carcinogenesis and (3) the creation of a novel screening approach to identify phospho-specific protein-protein interactions and E3 ubiquitin ligase substrates in a genome-wide manner. Since downstream targets are likely to be misregulated in cells lacking normal BRCA1 function, our screen is expected to reveal novel targets for drug development and novel biomarkers for assessing cellular responses to genotoxic stress.

描述（由申请人提供）：在日益工业化的环境中，我们越来越多地接触各种外源化合物，其中许多是 DNA 损伤剂，导致癌症发病率增加。提高我们对细胞如何响应和修复 DNA 损伤的理解将使我们能够更好地评估暴露于环境因素时的基因毒性应激，并开发新的分子策略来改善细胞对 DNA 损伤的反应。在本提案中，我们重点关注乳腺癌-1易感基因产物（BRCA1）（一种参与DNA损伤信号转导的关键分子）负责乳腺癌和卵巢特异性肿瘤抑制的机制。我们之前将 BRCA1 的 BRCT 结构域鉴定为磷酸丝氨酸/苏氨酸结合结构域，并且这种磷酸结合功能最近被证明对于 BRCA1 的肿瘤抑制功能至关重要。我们将进行全面的蛋白质组筛选，以确定 BRCA1 的直接 BRCT 结构域依赖性结合伴侣，以及雌激素反应组织中 BRCA1 与其结合伴侣 BARD1 复合的泛素连接酶活性的下游靶标。拟议的研究非常符合我们实验室的长期目标，即从分子细节上了解细胞信号通路的翻译后修饰如何与模块化结合域一起调节细胞对 DNA 损伤和细胞周期进展的反应的多个方面。筛选结果应提供 (1) 对雌激素反应性乳腺细胞中所有 BRCA1 相互作用蛋白进行全面、公正的鉴定，并确定它们是否是 BRCA1-BARD1 泛素连接酶活性的底物； (2) 鉴定 DNA 损伤诱发致癌风险增加的潜在靶点和生物标志物，以及 (3) 创建一种新的筛选方法，以全基因组方式鉴定磷酸特异性蛋白质-蛋白质相互作用和 E3 泛素连接酶底物。由于下游靶点在缺乏正常 BRCA1 功能的细胞中可能会被错误调节，因此我们的筛选有望揭示药物开发的新靶点和用于评估细胞对基因毒性应激反应的新生物标志物。