Faas F G A, Rieger B, van Vliet L J, Cherny D I
Department of Imaging Science and Technology Delft University of Technology, Delft, The Netherlands.
Biophys J. 2009 Aug 19;97(4):1148-57. doi: 10.1016/j.bpj.2009.06.015.
DNA is a very important cell structural element, which determines the level of expression of genes by virtue of its interaction with regulatory proteins. We use electron (EM) and atomic force microscopy (AFM) to characterize the flexibility of double-stranded DNA ( approximately 150-950 nm long) close to a charged surface. Automated procedures for the extraction of DNA contours ( approximately 10-120 nm for EM data and approximately 10-300 nm for AFM data) combined with new statistical chain descriptors indicate a uniquely two-dimensional equilibration of the molecules on the substrate surface regardless of the procedure of molecule mounting. However, in contrast to AFM, the EM mounting leads to a noticeable decrease in DNA persistence length together with decreased kurtosis. Analysis of local bending on short length scales (down to 6 nm in the EM study) shows that DNA flexibility behaves as predicted by the wormlike chain model. We therefore argue that adhesion of DNA to a charged surface may lead to additional static bending (kinking) of approximately 5 degrees per dinucleotide step without impairing the dynamic behavior of the DNA backbone. Implications of this finding are discussed.
DNA是一种非常重要的细胞结构元件,它通过与调节蛋白的相互作用来决定基因的表达水平。我们使用电子显微镜(EM)和原子力显微镜(AFM)来表征靠近带电表面的双链DNA(长度约为150 - 950纳米)的柔韧性。结合新的统计链描述符的DNA轮廓提取自动化程序(EM数据约为10 - 120纳米,AFM数据约为10 - 300纳米)表明,无论分子固定程序如何,分子在底物表面都具有独特的二维平衡。然而,与AFM不同的是,EM固定会导致DNA持续长度显著降低,同时峰度也降低。对短长度尺度(EM研究中低至6纳米)的局部弯曲分析表明,DNA的柔韧性符合蠕虫状链模型的预测。因此,我们认为DNA与带电表面的粘附可能会导致每二核苷酸步长额外产生约5度的静态弯曲(扭结),而不会损害DNA主链的动态行为。本文讨论了这一发现的意义。