Stacking-induced fluorescence increase reveals allosteric interactions through DNA.

Stacking-induced fluorescence increase reveals allosteric interactions through DNA.
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DOI:
10.1093/nar/gky887
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发表时间:
2018-11-30
影响因子:
14.9
通讯作者:
Magennis SW
Magennis SW
中科院分区:
生物学2区
文献类型:
--
作者:
Morten MJ;Lopez SG;Steinmark IE;Rafferty A;Magennis SW

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从基因表达到纳米技术,理解和控制DNA需要对其高阶结构和动力学有详细的了解。在这里,我们利用环境敏感的花青素染料的光异构化来探测DNA结构的局部和全局变化。我们报道了一种共价连接的Cy3染料在双链DNA的缺口、间隙或悬垂区堆叠时,荧光强度和寿命都有很强的增强。这被用来探测DNA发夹的杂交动力学,直到单分子水平。我们还表明,改变单个碱基位点的位置最多20个碱基对调节染料- dna相互作用,表明通过主干变构相互作用。叠致荧光增强(SIFI)现象在核酸的结构、动力学和反应性研究中应得到广泛应用。
From gene expression to nanotechnology, understanding and controlling DNA requires a detailed knowledge of its higher order structure and dynamics. Here we take advantage of the environment-sensitive photoisomerization of cyanine dyes to probe local and global changes in DNA structure. We report that a covalently attached Cy3 dye undergoes strong enhancement of fluorescence intensity and lifetime when stacked in a nick, gap or overhang region in duplex DNA. This is used to probe hybridization dynamics of a DNA hairpin down to the single-molecule level. We also show that varying the position of a single abasic site up to 20 base pairs away modulates the dye–DNA interaction, indicative of through-backbone allosteric interactions. The phenomenon of stacking-induced fluorescence increase (SIFI) should find widespread use in the study of the structure, dynamics and reactivity of nucleic acids.
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