Elucidating mechanisms underlying replication checkpoint control

阐明复制检查点控制的底层机制

基本信息

批准号：
10620981
负责人：
Junjie Chen
金额：
$ 49.94万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-16 至 2028-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10620981
关键词：
Address Antineoplastic Agents Binding CHEK1 gene Cell Cycle Cell Cycle Checkpoint Cell Cycle Progression Cell Cycle Regulation Cell Line Cell Proliferation Cell Survival Cells Chemotherapy and/or radiation Clinical Trials DNA DNA Damage DNA Repair DNA biosynthesis DNA lesion DNA replication fork Defect ETAA1 Ensure Event Genetically Engineered Mouse Genome Goals Immune checkpoint inhibitor Lead Lesion MCM2 gene Maintenance Mediating Mitosis Molecular Monitor Oncogenes Pathway interactions Phosphorylation Phosphotransferases Play Polymerase Process Proliferating Protein Kinase Proteins Radiation Radiation Induced DNA Damage Regulation Replication Initiation Role S phase SS DNA BP Signal Transduction Single-Stranded DNA Structure System TOPBP1 Gene Testing Work cancer therapy chemotherapeutic agent established cell line genome integrity helicase inhibitor prevent protein degradation protein function recruit replication stress response upstream kinase

项目摘要

PROJECT SUMMARY The maintenance of genomic integrity after DNA damage and replication stress depends on the coordination of DNA repair and cell cycle checkpoints. The replication checkpoint pathway, which comprises two critical protein kinases, ATR and CHK1, has an essential role in this coordination. Inhibition of the replication checkpoint results in cell lethality in response to replication stress induced by oncogenes, radiation, or chemotherapeutic agents. Indeed, several replication checkpoint inhibitors are being tested in clinical trials as potential anticancer agents. Our objective in this application is to provide a detailed mechanistic understanding of replication checkpoint control, which may help us develop the best strategies for using these checkpoint inhibitors in cancer therapy. We have studied replication checkpoint control for many years. We demonstrated that the ATR-dependent replication checkpoint can be activated by a variety of DNA lesions. In addition, ATR not only activates CHK1 but also phosphorylates many other substrates, including MCM, H2AX, and others at or near stalled replication forks to initiate replication checkpoint signaling. Moreover, we have identified and studied several key proteins such as TOPBP1, ETAA1, and Claspin involved in replication checkpoint control. Many key proteins involved in DNA replication and replication checkpoint control are essential for cell survival. Thus, it remains challenging to fully understand the roles of DNA replication and replication checkpoint proteins in cell cycle progression. Toward this end, we recently took advantage of the dTAG- mediated protein degradation system and established cell lines with inducible degradation of several essential DNA replication and replication checkpoint proteins. Initial analyses of events following the depletion of these essential proteins revealed some unexpected observations, which led us to re-evaluate replication checkpoint control and the mechanisms underlying cell cycle transitions. In this project, we will determine the essential functions of these DNA replication and checkpoint proteins in S phase and cell cycle progression. We anticipate that results from these studies will provide a better understanding of replication checkpoint control, especially how DNA replication and replication checkpoint are coordinated to ensure S phase progression and cell survival.

项目概要 DNA 损伤和复制应激后基因组完整性的维持取决于 DNA 修复和细胞周期检查点。复制检查点途径，包括两个关键的蛋白激酶 ATR 和 CHK1 在这种协调中发挥着重要作用。复制抑制检查点导致细胞因癌基因、辐射或诱导的复制应激而致死化疗剂。事实上，几种复制检查点抑制剂正在临床试验中进行测试：潜在的抗癌剂。我们在此应用程序中的目标是提供详细的机制理解复制检查点控制，这可以帮助我们制定使用这些检查点的最佳策略癌症治疗中的抑制剂。我们研究复制检查点控制很多年了。我们证明了 ATR 依赖性复制检查点可以被多种DNA损伤激活。此外，ATR不仅激活CHK1 而且还磷酸化许多其他底物，包括 MCM、H2AX 和其他复制处于或接近停滞状态的底物分叉以启动复制检查点信号。此外，我们还鉴定并研究了几种关键蛋白质例如参与复制检查点控制的 TOPBP1、ETAA1 和 Claspin。许多参与 DNA 复制和复制检查点控制的关键蛋白质对于细胞至关重要生存。因此，充分理解 DNA 复制和复制的作用仍然具有挑战性。细胞周期进展中的检查点蛋白。为此，我们最近利用了 dTAG- 介导的蛋白质降解系统并建立了具有几种必需物质诱导降解的细胞系 DNA 复制和复制检查点蛋白。对这些资源耗尽后事件的初步分析必需蛋白质揭示了一些意想不到的观察结果，这导致我们重新评估复制检查点细胞周期转变的控制和机制。在这个项目中，我们将确定这些 DNA 复制和检查点蛋白的基本功能 S期和细胞周期进程。我们预计这些研究的结果将提供更好的结果了解复制检查点控制，尤其是 DNA 复制和复制检查点是如何进行的协调以确保 S 期进展和细胞存活。