Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles.

Bending modes of DNA directly addressed by cryo-electron microscopy of DNA minicircles.
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DOI:
10.1093/nar/gkp137
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发表时间:
2009-05
影响因子:
14.9
通讯作者:
Stasiak A
Stasiak A
中科院分区:
生物学2区
文献类型:
--
作者:
Demurtas D;Amzallag A;Rawdon EJ;Maddocks JH;Dubochet J;Stasiak A

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我们使用低温电子显微镜(cro - em)来研究94 bp长的DNA小圆的三维形状,并解决这样的短DNA分子的环化是否需要在DNA中形成尖锐的局部扭结,或者是否必要的弯曲可以在DNA弹性的标准模型的限制内重新分布和完成。通过比较共价闭合、有缺口和有缺口的DNA小环的形状,我们得出结论,94 bp长的共价闭合和有缺口的DNA小环不显示尖锐的扭结,而含有非常灵活的单链区域的有缺口的DNA分子确实显示尖锐的扭结。我们用Bal31核酸酶在94 bp长的小圆中探测扭结的存在,证实了冷冻电镜研究的结果。
We use cryo-electron microscopy (cryo-EM) to study the 3D shapes of 94-bp-long DNA minicircles and address the question of whether cyclization of such short DNA molecules necessitates the formation of sharp, localized kinks in DNA or whether the necessary bending can be redistributed and accomplished within the limits of the elastic, standard model of DNA flexibility. By comparing the shapes of covalently closed, nicked and gapped DNA minicircles, we conclude that 94-bp-long covalently closed and nicked DNA minicircles do not show sharp kinks while gapped DNA molecules, containing very flexible single-stranded regions, do show sharp kinks. We corroborate the results of cryo-EM studies by using Bal31 nuclease to probe for the existence of kinks in 94-bp-long minicircles.
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发表时间: 1996-07-12
影响因子: 5.6
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