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含有N(4)C-乙烯-N(4)C跨链交叉链的DNA复合体的合成和表征

D M Noll¹, A M Noronha, P S Miller

¹Department of Biochemistry and Molecular Biology, School of Hygiene and Public Health, Johns Hopkins University, 615 North Wolfe Street, Baltimore, MD 21205, USA.

Journal of the American Chemical Society

|July 27, 2001

概括

研究人员合成了具有特定跨链跨链的DNA复合体,为研究DNA修复和生物物理实验提供了新的工具.

科学领域:

分子生物学分子生物学
有机化学有机化学
生物化学生物化学

背景情况:

DNA 链间交叉链 (ICL) 是关键的 DNA 损伤病变.
了解ICL的结构和功能后果对于DNA修复研究至关重要.

研究的目的:

开发一种合成含有定义的N(4)C-乙烯-N(4)C跨链交联 (C-C) 的DNA复合体的方法.
为生物物理和生物化学研究创建基质,包括DNA修复机制.

主要方法:

在受控孔玻璃支上合成DNA复合体,采用可转换核酸方法.
引入C-C交叉链路,通过将三醇组被氨基乙基部分所取代.
通过使用胺酸化学方法从交叉连接中延长两个DNA链.
通过高性能液体染色学 (HPLC) 进行净化,并通过酶性消化和MALDI-TOF质谱学进行表征.

主要成果:

成功合成了具有不同长度的 A/T 基对绕着 C-C 交叉链接的 DNA 复合体.
观察到A/T基对双重组的合作性变性,其中有三个或四个侧面对.
证明T4DNA结合酶可以结合这些交叉链接的复合体,即使交叉链接靠近结合部位.

结论:

开发的合成策略允许精确地放置C-C跨链交叉链接在DNA复合体中.
这些交叉连接的双重体是结反应的合适基质,表明它们在构建更复杂的DNA结构中的实用性.
合成的DNA结构为研究DNA修复途径和其他与DNA相关的生物化学过程提供了宝贵的工具.

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