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

Extreme bendability of DNA less than 100 base pairs long revealed by single-molecule cyclization.
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DOI:
10.1126/science.1224139
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发表时间:
2012-08-31
期刊:
Science (New York, N.Y.)
影响因子:
--
通讯作者:
Ha T
Ha T
中科院分区:
其他
文献类型:
--
作者:
Vafabakhsh R;Ha T

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DNA的经典观点假定DNA在低于持久长度(<150 bp)时必须是刚性的,但最近针对这一点的研究产生了矛盾的结果。我们开发了一种基于荧光的、无蛋白质的测定方法,用于真实的研究单个DNA分子的环化。该测定对急剧弯曲的瞬时物质的平衡群体进行采样,该平衡群体在单分子机械测量中被完全抑制,并且比传统的基于连接酶的测定中采样的连接物质在生物学上更相关。成环率在67 ~ 106 bp之间具有明显的弱的长度依赖性,不能用蠕虫状链模型来描述。许多生物学上重要的蛋白质-DNA相互作用,涉及低于100 bp的DNA的成环和弯曲,可能使用DNA的这种内在的弯曲性。
The classical view of DNA posits that DNA must be stiff below the persistence length (<150 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 which is entirely suppressed in single molecule mechanical measurements and is biologically more relevant than the ligated species sampled in the traditional ligase-based assay. The looping rate has a remarkably weak length dependence between 67 and 106 bp that can not be described by the worm-like chain model. Many biologically significant protein-DNA interactions that involve looping and bending of DNA below 100 bp likely use this intrinsic bendability of DNA.
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