COBRE: UNE MED CTR: P8:MRN AND RPA PROTEIN-PROTEIN INTERACTIONS IN DNA DAMAGE

COBRE：UNE MED CTR：P8：MRN 和 RPA 蛋白质-蛋白质相互作用在 DNA 损伤中的作用

• 批准号: 7382062
• 负责人: Gregory G Oakley
• 金额: $ 24.93万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-07-01 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7382062
• 关键词: COBRE; UNE; MED; CTR; P8

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The long-term goal of the proposed research is to understand how genomic integrity is preserved in cells. Highly complex surveillance mechanisms, consisting of DMA repair, DMA replication and checkpoint proteins, have developed to maintain genomic integrity. Dysfunction of these mechanisms can lead to a variety of clinical symptoms including an increased risk of cancer. Progress in our understanding of the downstream effectors proteins activated in response to DMA damage that lead to cell cycle arrest and DNA repair have been made but the mechanisms by which DNA damage is detected and signaled remain elusive. Therefore, we aim to define the sensor components of the replication stress response pathway and determine how they work cooperatively in stabilizing stalled replication forks during S-phase of the cell cycle. Among the proteins thought to be involved in this response include the MRN complex (composed of MRE11, RAD50 and NBS1), and RPA, (RPA, the major single-stranded DNA binding protein in eukaryotic cells composed of three subunits p70, p34, p14), proteins that are intricately involved in DNA metabolism and maintenance of genomic stability. We have recently identified a protein/protein interaction between the MRN complex and RPA. We believe that the MRN complex and RPA work together in response to DNA damage and stalled replication forks. However, how the MRN complex and RPA sense and initiate a response to stalled replication forks remains undefined. This proposal will test the hypothesis that stalled replication forks stimulate the MRN complex in an RPA-dependent manner to stabilize and repair damage at all sites of stalled replication. To test this hypothesis the following aims are proposed: Aim 1: To characterize the direct protein-protein interactions of RPA and the MRN complex. Aim 2: Define the role of RPA in the recruitment of the M/R/N complex to sites of stalled replication. Aim 3: Characterize the role of RPA and phosphorylation in the tethering of DNA by the MRN complex.

该子项目是利用 NIH/NCRR 资助的中心拨款提供的资源的众多研究子项目之一。子项目和研究者 (PI) 可能已从另一个 NIH 来源获得主要资金，因此可以在其他 CRISP 条目中得到体现。列出的机构是中心的机构，不一定是研究者的机构。拟议研究的长期目标是了解细胞中如何保持基因组完整性。由 DMA 修复、DMA 复制和检查点蛋白组成的高度复杂的监视机制已经开发出来，以维持基因组完整性。这些机制的功能障碍可能导致各种临床症状，包括癌症风险增加。我们对响应 DMA 损伤而激活的下游效应蛋白的理解已经取得进展，这些蛋白会导致细胞周期停滞和 DNA 修复，但检测和发出信号的 DNA 损伤的机制仍然难以捉摸。因此，我们的目标是定义复制应激反应途径的传感器组件，并确定它们如何在细胞周期 S 期期间协同工作来稳定停滞的复制叉。 参与这种反应的蛋白质包括 MRN 复合物（由 MRE11、RAD50 和 NBS1 组成）和 RPA（RPA，真核细胞中主要的单链 DNA 结合蛋白，由三个亚基 p70、p34、p14 组成），这些蛋白质复杂地参与 DNA 代谢和基因组稳定性的维持。我们最近发现了 MRN 复合物和 RPA 之间的蛋白质/蛋白质相互作用。我们相信 MRN 复合物和 RPA 共同作用来应对 DNA 损伤和停滞的复制叉。然而，MRN 复合体和 RPA 如何感知并启动对停滞的复制叉的响应仍然未定义。该提案将测试以下假设：停滞的复制叉以 RPA 依赖性方式刺激 MRN 复合物，以稳定和修复所有停滞复制位点的损伤。为了检验这一假设，提出了以下目标： 目标 1：表征 RPA 和 MRN 复合物的直接蛋白质-蛋白质相互作用。目标 2：定义 RPA 在将 M/R/N 复合物募集到复制停滞位点中的作用。目标 3：表征 RPA 和磷酸化在 MRN 复合物束缚 DNA 中的作用。