Probing the elastic limit of DNA bending.

Probing the elastic limit of DNA bending.
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DOI:
10.1093/nar/gku735
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发表时间:
2014
影响因子:
14.9
通讯作者:
Kim HD
Kim HD
中科院分区:
生物学2区
文献类型:
--
作者:
Le TT;Kim HD

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双链DNA (dsDNA)的急剧弯曲在基因组的结构和功能中起着至关重要的作用。然而,dsDNA弯曲的弹性极限仍然存在争议。在这里,我们使用单分子荧光共振能量转移测量了轮廓长度在40到200 bp之间的小dsDNA环的打开率。环路寿命与环路尺寸的关系揭示了弯曲应力的临界转变。在临界环尺寸以上,环寿命随环尺寸变化的方式与弹性弯曲应力一致,但低于临界环尺寸,对环尺寸变得不那么敏感,表明dsDNA软化。添加5 mM镁后,临界环尺寸从~ 60 bp增加到~ 100 bp。我们表明,我们的结果与可卷曲的蠕虫状链模型在定量上是一致的,而且,可以再现先前报道的dsDNA在50到200 bp范围内的环概率。我们的发现对急剧弯曲的dsDNA的能量学有了新的认识。
Sharp bending of double-stranded DNA (dsDNA) plays an essential role in genome structure and function. However, the elastic limit of dsDNA bending remains controversial. Here, we measured the opening rates of small dsDNA loops with contour lengths ranging between 40 and 200 bp using single-molecule Fluorescence Resonance Energy Transfer. The relationship of loop lifetime to loop size revealed a critical transition in bending stress. Above the critical loop size, the loop lifetime changed with loop size in a manner consistent with elastic bending stress, but below it, became less sensitive to loop size, indicative of softened dsDNA. The critical loop size increased from ∼60 bp to ∼100 bp with the addition of 5 mM magnesium. We show that our result is in quantitative agreement with the kinkable worm-like chain model, and furthermore, can reproduce previously reported looping probabilities of dsDNA over the range between 50 and 200 bp. Our findings shed new light on the energetics of sharply bent dsDNA.
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