Chemical and Biological Engineering, Koc University, Sariyer, Istanbul, Turkey.
PLoS One. 2012;7(8):e44392. doi: 10.1371/journal.pone.0044392. Epub 2012 Aug 29.
The cyclobutane pyrimidine dimer (CPD) and 6-4 lesion formations along with the specific breaks on strands are the most common type of DNA damage caused by Ultraviolet light (UV) irradiation. CPD photolyase I and II construct two subfamilies of flavoproteins, which have recognition and repair capabilities of CPD sites on both single stranded (ssDNA) and double stranded DNA (dsDNA) with the aid of blue light energy. The other types of flavoprotein family consist of cryptochromes (CRY) that act as photoreceptors in plants, or circadian rhythm regulators in animals. Recent findings showed that a specific type of Cryptochrome-Drosophila, Arabidopsis, Synechocystis, Human (CRY-DASH) has photorepair activity on ssDNA. In this work, real-time interactions between CRY-DASH and ss/dsDNA as well as the interactions between Vibrio cholerae photolyase (VcPHR) and ss/dsDNA were investigated using Surface Plasmon Resonance (SPR). The interactions were then characterized and compared in order to investigate the effect of different types of flavoprotein on UV damaged ss/dsDNA. SPR results confirm the specific binding of VcPHR and CRY-DASH with UV treated DNA. This study is the first instance to quantify the interactions of UV treated and untreated DNA with flavoproteins.
环丁烷嘧啶二聚体（CPD）和 6-4 损伤以及链上的特定断裂是紫外线（UV）照射引起的最常见的 DNA 损伤类型。CPD 光解酶 I 和 II 构建了两个黄素蛋白亚家族，它们在蓝光能量的帮助下具有识别和修复单链（ssDNA）和双链 DNA（dsDNA）上 CPD 位点的能力。其他类型的黄素蛋白家族包括在植物中充当光受体或在动物中充当昼夜节律调节剂的隐花色素（CRY）。最近的研究结果表明，一种特定类型的隐花色素-Drosophila、拟南芥、集胞藻、人类（CRY-DASH）对 ssDNA 具有光修复活性。在这项工作中，使用表面等离子体共振（SPR）研究了 CRY-DASH 与 ss/dsDNA 之间的实时相互作用以及霍乱弧菌光解酶（VcPHR）与 ss/dsDNA 之间的相互作用。然后对相互作用进行了表征和比较，以研究不同类型的黄素蛋白对 UV 损伤的 ss/dsDNA 的影响。SPR 结果证实了 VcPHR 和 CRY-DASH 与经 UV 处理的 DNA 的特异性结合。这项研究首次定量研究了经 UV 处理和未经处理的 DNA 与黄素蛋白的相互作用。
