Molecular Mechanism of Mammalian DNA Excision Repair, DNA Damage Checkpoints and the Circadian Clock

 描述（由申请人提供）：我们研究三个相互关联的主题：哺乳动物DNA切除修复，DNA损伤检查点和生物钟。我们最近在这三个领域都取得了一些发现，加深了对这三个系统之间相互联系的理解，并与人类健康和疾病治疗直接相关。我们开发了XR-seq（切除修复测序）方法，以单核苷酸分辨率生成整个人类基因组的切除修复图谱;我们重建了一个将核苷酸切除修复与DNA损伤检查点联系起来的体外系统;我们发现了生物钟对切除修复的调节，并开发了新的方法来了解哺乳动物生物钟的分子机制。我们将把这些新的概念和技术进步应用于以下目标：（1）了解影响致癌剂和抗癌剂引起的DNA损伤的切除修复的因素，（2）改进切除修复-检查点偶联的体外系统以用于抗癌药物筛选和验证，以及（3）表征哺乳动物生物钟的分子机制，并基于人类DNA切除修复活性的时钟控制开发癌症预防和治疗策略。这种方法是创新的，因为它结合了生物化学/生物物理学，细胞生物学，遗传学和动物生理学，以了解哺乳动物DNA切除修复，DNA损伤检查点和生物钟之间的机制基础和联系。这项拟议中的研究意义重大，因为它与癌症预防和治疗有关。

 DESCRIPTION (provided by applicant): We work on three inter-related subjects: mammalian DNA excision repair, DNA damage checkpoints, and the circadian clock. We have made a number of recent discoveries in all three areas that lead to deeper understanding of the interconnections between these three systems and are directly relevant to human health and disease treatment. We developed the XR-seq (excision Repair-sequencing) method to generate an excision repair map of the entire human genome at single nucleotide resolution; we have reconstituted an in vitro system that links nucleotide excision repair to the DNA damage checkpoint; and we have discovered regulation of excision repair by the circadian clock and developed novel approaches to understand the molecular mechanism of the mammalian circadian clock. We will apply these new conceptual and technical advances for the following objectives: (1) understand factors that affect excision repair of DNA damage by carcinogenic and by anticancer agents, (2) improve the in vitro system of excision repair-checkpoint coupling for potential use in anticancer drug screening and validation, and (3) characterize the molecular mechanism of the mammalian circadian clock and develop strategies of cancer prevention and cancer treatment based on clock control of human DNA excision repair activity. This approach is innovative because it combines biochemistry/biophysics, cell biology, genetics and animal physiology to understand the mechanistic basis and connections between mammalian DNA excision repair, DNA damage checkpoints, and of the circadian clock. The proposed research is significant because of its relevance to cancer prevention and treatment.