Photochemical determination of different DNA structures
Photochemical determination of different DNA structures
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DOI：
10.1038/nprot.2006.467
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Sugiyama, Hiroshi
中科院分区：
文献类型：
--
作者：
Xu, Yan;Tashiro, Ryu;Sugiyama, Hiroshi
DNA的各种构象被认为具有重要的生物学作用。因此，研究与生物事件相关的局部DNA构象变化对于理解DNA的功能是必不可少的。我们报道了5-氟尿嘧啶在五种局部DNA结构中的光反应性：A、B和Z形式，蛋白质诱导的DNA扭结和G-四链形式。这些研究证明了DNA的局部结构与产生的尿嘧啶-5-基自由基光诱导的光化学产物之间的详细关系，并表明这种光化学方法可以用来检测DNA结构。在这里，我们详细描述了我们实验室开发的通过对含有5-卤尿嘧啶的DNA进行光辐照产物分析来探测DNA构象的程序。该方案包括含5-氟尿嘧啶的DNA的制备和光产物的表征，可在2周内完成。
The various conformations of DNA are thought to have important biological roles. Investigation of the local DNA conformational changes associated with biological events is therefore essential to an understanding of the functions of DNA. We have reported the photoreactivities of 5-halouracil in the five characteristic local DNA structures: the A, B and Z forms, protein-induced DNA kinks and the G-quadruplex form. These studies demonstrate the detailed relationships between the local DNA structures and the photochemical products of photoinduced hydrogen abstraction by the resulting uracil-5-yl radicals, and show that this photochemical method can be used to detect DNA structures. Here, we describe in detail procedures that have been developed in our laboratory for probing DNA conformations by product analysis of photoirradiated 5-halouracil-containing DNA. The protocol includes the preparation of 5-halouracil-containing DNA and the characterization of the photoproducts, and it can be completed in 2 weeks.