Understanding the mechanobiology of DNA-protein interactions at the single-molecule level

DNA-蛋白质相互作用是DNA复制、转录和修复等基本细胞过程的基础，参与这些过程的蛋白质是各种癌症治疗的靶点。许多受损的DNA-蛋白质复合体的结构显示出DNA底物的赤裸裸的弯曲，这表明蛋白质可能在识别损伤位置的过程中对DNA施加力来弯曲它，或者为了促进修复。两个这样的蛋白质是FEN1(对NER和Okasaki片段处理必不可少)和XPA(对NER必不可少)。在这里，我们将开发新的仪器来研究力如何在基础水平上调节DNA修复中的DNA-蛋白质相互作用，并建立高通量的筛选方法，模拟DNA底物上的生理力(由于转录和包装而自然出现在细胞中)，以改进修复抑制剂的HIT鉴定。

DNA-protein interactions underpin essential cellular processes such as DNA replication, transcription, and repair, and proteins involved in these processes are targeted in a variety of cancer therapies. Many structures of damaged DNA-protein complexes reveal stark bends in the DNA substrate, suggesting that proteins may exert force on the DNA to bend it during recognition of the damage site, or for catalytic enhancement of the repair. Two such proteins are FEN1 (essential for NER and Okasaki fragment processing) and XPA (essential to NER). Here we will develop novel instrumentation to investigate how force modulates the DNA-protein interactions involved in DNA repair at a fundamental level, and establish high-throughput screening methods which mimic physiological forces on DNA substrates (which naturally arise in cells due to transcription and packaging) to improve hit identification for repair inhibitors.