Visualizing the Path of DNA through Proteins Using DREEM Imaging

Molecular Cell
Dong WuD A Erie

Abstract

Many cellular functions require the assembly of multiprotein-DNA complexes. A growing area of structural biology aims to characterize these dynamic structures by combining atomic-resolution crystal structures with lower-resolution data from techniques that provide distributions of species, such as small-angle X-ray scattering, electron microscopy, and atomic force microscopy (AFM). A significant limitation in these combinatorial methods is localization of the DNA within the multiprotein complex. Here, we combine AFM with an electrostatic force microscopy (EFM) method to develop an exquisitely sensitive dual-resonance-frequency-enhanced EFM (DREEM) capable of resolving DNA within protein-DNA complexes. Imaging of nucleosomes and DNA mismatch repair complexes demonstrates that DREEM can reveal both the path of the DNA wrapping around histones and the path of DNA as it passes through both single proteins and multiprotein complexes. Finally, DREEM imaging requires only minor modifications of many existing commercial AFMs, making the technique readily available.

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Citations

Jan 18, 2016·Molecular Cell·Delphine Benarroch-PopivkerMarie-Josèphe Giraud-Panis
Apr 5, 2017·Annual Review of Biophysics·Stephan Uphoff, David J Sherratt
Jun 5, 2018·Journal of Molecular Recognition : JMR·Samrat DuttaMartin Guthold
Feb 17, 2017·The Journal of Biological Chemistry·Nicholas L AdkinsCraig L Peterson
Jun 27, 2020·Proceedings of the National Academy of Sciences of the United States of America·Kira C BradfordDorothy A Erie
Jul 5, 2017·Seminars in Cell & Developmental Biology·Emily C BeckwittBennett Van Houten

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