DNA和DNA修复蛋白的原子力显微镜成像:在放射生物学研究中的应用。

D Pang, B Vidic, J Rodgers, B L Berman, A Dritschilo
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引用次数: 15

摘要

利用原子力显微镜(AFM),可以在纳米分辨率下对生物标本的三维结构进行成像。此外,样品可以在空气或流体环境中成像。AFM操作成像的敲击模式在可视化软生物结构(如DNA、蛋白质和膜)方面取得了重大进展。在这里,我们回顾了该仪器在辐射生物学研究中的应用原理。我们重点介绍了参与损伤DNA修复过程的蛋白质,包括聚(adp -核糖)聚合酶、Ku蛋白和DNA蛋白激酶。对DNA损伤和修复特征的新观察已经通过DNA和蛋白质-DNA相互作用的直接可视化来解决,例如观察到Ku蛋白能够在体外物理连接DNA片段。AFM为研究与DNA损伤和修复过程相关的生物学上重要的分子相互作用提供了一个强大的工具。
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Atomic force microscope imaging of DNA and DNA repair proteins: applications in radiobiological research.

By using the atomic force microscope (AFM), three-dimensional structures of biological specimens may be imaged at nanometer resolution. Furthermore, samples can be imaged in air or in fluid environments. The tapping mode of AFM operation for imaging has offered a significant advance in visualizing soft biological structures, such as DNA, proteins, and membranes. Here, we review the principles underlying the application of this instrument to radiation biological investigations. We focus on examples of proteins involved in the processes of repair of damaged DNA, including poly(ADP-ribose) polymerase, Ku protein, and DNA protein kinase. Novel observations on the character of DNA damage and repair have been addressed by direct visualization of DNA and protein-DNA interactions, such as the observation that the Ku protein is capable of physically joining DNA fragments in vitro. The AFM offers a powerful tool for investigating biologically important molecular interactions that are relevant to DNA damage and repair processes.

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