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Study of the interaction of DNA and histones by spin-stretching and droplet evaporation
Authors:YuYing Liu  PengYe Wang  ShuoXing Dou  WeiWei Zhang  XueJin Wang  HongYi Sang
Affiliation:(1) Department of Chemistry and Chemical Engineering, Polytechnic University, Brooklyn, NY 11201, USA;(2) Department of Cell Biology and Physiology, University of Pittsburgh School of Medicine, Hillman Cancer Center, UPCI Research Pavilion, Pittsburgh, PA 15213, USA
Abstract:
Molecular combing is a powerful and simple method for aligning DNA molecules onto a surface. Using this technique combined with fluorescence microscopy, DNA-histone complexes are stretched on a hydrophobic polymethyl methacrylate (PMMA) surface and observed directly. We have developed a new method to stretch single DNA-histone complexes, termed spin-stretching. The results show that the histones markedly enhance DNA binding to the PMMA surface. DNA winds around the histones and therefore decreases in length. The number of histones that bind to each DNA molecule is found to correlate with the histone concentration. The combed DNA-histone complexes are found to depend on two factors: the binding force on the surface and the centrifugal force at its local position. Na+ ions should compete with histones for binding to DNA; however, the observed competitive binding effect of Na+ ions at low concentrations was negligible.
Keywords:molecular combing   histone   DNA-histone complex   spin-stretching   droplet evaporation
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