How does temperature impact the conformation of single DNA molecules below melting temperature?
How does temperature impact the conformation of single DNA molecules below melting temperature?
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DOI：
10.1093/nar/gkx1285
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发表时间：
2018-02-28
影响因子：
14.9
通讯作者：
Tardin C
中科院分区：
文献类型：
--
作者：
Brunet A;Salomé L;Rousseau P;Destainville N;Manghi M;Tardin C
双链DNA分子在生物过程中经历剧烈的结构变化，例如转录，在转录期间，它在RNA聚合酶的作用下局部打开。不同的生物物理学研究支持了这一观点，发现具有不同序列的DNA分子的灵活性作为温度的函数意外增加，这与越来越多的局部变性序列的形成是一致的。在这里，我们利用我们的能力来检测DNA上发生的细微变化，通过使用高通量拴系粒子运动来质疑生理盐条件下双链DNA中气泡的存在，通过它们对DNA分子的构象影响，范围从几百到几千个碱基对。我们的结果与以前发表的结果有显著的不同，因为我们没有检测到任何意想不到的变化，DNA的灵活性低于熔解温度。相反，我们测量的弯曲模量，保持稳定的温度，如预期的完整的双链DNA。
The double stranded DNA molecule undergoes drastic structural changes during biological processes such as transcription during which it opens locally under the action of RNA polymerases. Local spontaneous denaturation could contribute to this mechanism by promoting it. Supporting this idea, different biophysical studies have found an unexpected increase in the flexibility of DNA molecules with various sequences as a function of the temperature, which would be consistent with the formation of a growing number of locally denatured sequences. Here, we take advantage of our capacity to detect subtle changes occurring on DNA by using high throughput tethered particle motion to question the existence of bubbles in double stranded DNA under physiological salt conditions through their conformational impact on DNA molecules ranging from several hundreds to thousands of base pairs. Our results strikingly differ from previously published ones, as we do not detect any unexpected change in DNA flexibility below melting temperature. Instead, we measure a bending modulus that remains stable with temperature as expected for intact double stranded DNA.
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