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锌离子及其他二价金属离子诱导双链DNA发生协同构象变化。

A cooperative conformational change in duplex DNA induced by Zn2+ and other divalent metal ions.

作者信息

Lee J S, Latimer L J, Reid R S

机构信息

Department of Biochemistry, University of Saskatchewan, Saskatoon, Canada.

出版信息

Biochem Cell Biol. 1993 Mar-Apr;71(3-4):162-8. doi: 10.1139/o93-026.

Abstract

Zn2+ and some other divalent metal ions bind to duplex DNA at pHs above 8 and cause a conformational change. This new structure does not bind ethidium, allowing the development of a rapid fluorescence assay. All duplex DNAs, regardless of sequence or G.C content, can form this structure. The rate of formation shows a strong dependence on temperature, pH, and Zn2+ concentration; at 20 degrees C, 1 mM Zn2+, and pH 8.6 the dismutation is half complete in 30 min. Addition of EDTA causes rapid reversion to 'B' DNA, showing that the new conformation retains two strands that are antiparallel. Unlike the ultraviolet or circular dichroism spectra, the nuclear magnetic resonance spectrum was informative since the imino protons of both A.T and G.C base pairs are lost upon addition of a stoichiometric amount of Zn2+. The pitch of the helix was estimated from gel electrophoresis of circular DNAs in the presence of Zn2+ and it contains at least 5% fewer base pairs per turn than 'B' DNA. The transformation is cooperative and shows hysteresis, suggesting that this is a distinct structure and not simply a minor variant of 'B' DNA. It is proposed to call this new structure 'M' DNA because of the intimate involvement of metal ions.

摘要

锌离子(Zn²⁺)和其他一些二价金属离子在pH值高于8时会与双链DNA结合,并引起构象变化。这种新结构不与溴化乙锭结合,从而可以开发一种快速荧光测定法。所有双链DNA,无论其序列或G.C含量如何,都能形成这种结构。形成速率强烈依赖于温度、pH值和Zn²⁺浓度;在20℃、1 mM Zn²⁺和pH 8.6的条件下,30分钟内歧化反应完成一半。加入乙二胺四乙酸(EDTA)会使DNA迅速恢复为“B”型,这表明新构象保留了两条反平行的链。与紫外光谱或圆二色光谱不同,核磁共振光谱提供了有用信息,因为加入化学计量的Zn²⁺后,A.T和G.C碱基对的亚氨基质子都会消失。通过在Zn²⁺存在下对环状DNA进行凝胶电泳来估计螺旋的螺距,结果表明其每圈碱基对比“B”型DNA至少少5%。这种转变具有协同性并表现出滞后现象,这表明这是一种独特的结构,而不仅仅是“B”型DNA的一个微小变体。由于金属离子的密切参与,建议将这种新结构称为“M”型DNA。

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