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单分子环化实验揭示了长度小于 100 个碱基对的 DNA 的极端柔韧性。

Extreme bendability of DNA less than 100 base pairs long revealed by single-molecule cyclization.

机构信息

Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

出版信息

Science. 2012 Aug 31;337(6098):1097-101. doi: 10.1126/science.1224139.

Abstract

The classical view of DNA posits that DNA must be stiff below the persistence length [<150 base pairs (bp)], but recent studies addressing this have yielded contradictory results. We developed a fluorescence-based, protein-free assay for studying the cyclization of single DNA molecules in real time. The assay samples the equilibrium population of a sharply bent, transient species that is entirely suppressed in single-molecule mechanical measurements and is biologically more relevant than the annealed species sampled in the traditional ligase-based assay. The looping rate has a weak length dependence between 67 and 106 bp that cannot be described by the worm-like chain model. Many biologically important protein-DNA interactions that involve looping and bending of DNA below 100 bp likely use this intrinsic bendability of DNA.

摘要

经典的 DNA 观点认为,DNA 在持久长度 [<150 个碱基对 (bp)] 以下必须是刚性的,但最近针对这一问题的研究得出了相互矛盾的结果。我们开发了一种基于荧光的、无蛋白质的测定方法,用于实时研究单链 DNA 分子的环化。该测定方法从一个急剧弯曲的、瞬时的物种的平衡种群中进行采样,而该物种在单分子力学测量中完全被抑制,并且比在传统的基于连接酶的测定中采样的退火物种更具有生物学相关性。环化率在 67 到 106 个碱基对之间具有较弱的长度依赖性,不能用蠕虫状链模型来描述。许多涉及 100bp 以下 DNA 环化和弯曲的重要生物蛋白-DNA 相互作用可能利用了 DNA 的这种固有柔韧性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7890/3565842/747707ccdc9b/nihms-437699-f0001.jpg

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