Molecular Studies Of Protein-DNA Interactions

利用各种生化、结构以及分子和细胞生物学方法，我们正在研究涉及DNA修复蛋白的各种细胞功能的分子机制。表面等离子体共振研究使我们能够实时监测在各种条件下在体外形成的关键蛋白质-蛋白质和蛋白质-DNA相互作用，以询问复合体的稳定性，并利用野生型和突变型蛋白质表征复合体形成的各个方面。荧光标记蛋白质和免疫荧光显微镜正被用来研究蛋白质之间的相互作用以及不同细胞间的运输。修复蛋白水平的调节、翻译后修饰和亚细胞定位正在被研究，以响应各种遗传毒性应激。使用原子力显微镜的单分子研究使我们能够可视化与DNA损伤结合的多蛋白质复合体。这些研究有助于阐明特定的DNA修复途径，以及可能与癌症和衰老相关的DNA损伤的一般细胞反应。

Using a variety of biochemical, structural, and molecular and cell biological approaches, we are studying the molecular mechanisms underlying the diverse array of cellular functions involving DNA repair proteins. Surface plasmon resonance studies allow us to monitor in real-time key protein-protein and protein-DNA interactions formed in vitro under a variety of conditions to ask about complex stability and to characterize various aspects of complex formation utilizing both wild type and mutant proteins. Fluorescently tagged proteins and immunofluorescence microscopy are being used to study protein-protein interactions as well as traffic within different cell compartments. Modulation of the levels of repair proteins, post-translational modifications, and subcellular localization are being examined in response to a variety of genotoxic stresses. Single molecule studies using atomic force microscopy allow us to visualize multi-protein complexes bound to DNA lesions. These studies can help shed light on specific DNA repair pathways as well as general cellular responses to DNA damage that may be relevant to cancer and aging.