Experimental maps of DNA structure at nucleotide resolution distinguish intrinsic from protein-induced DNA deformations
Azad, Robert N.
Sagendorf, Jared M.
Tullius, Thomas D.
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Citation (published version)Robert N Azad, Dana Zafiropoulos, Douglas Ober, Yining Jiang, Tsu-Pei Chiu, Jared M Sagendorf, Remo Rohs, Thomas D Tullius. 2018. "Experimental maps of DNA structure at nucleotide resolution distinguish intrinsic from protein-induced DNA deformations." Nucleic Acids Research, Volume 46, Issue 5, pp. 2636 - 2647 (12). https://doi.org/10.1093/nar/gky033
Recognition of DNA by proteins depends on DNA sequence and structure. Often unanswered is whether the structure of naked DNA persists in a protein–DNA complex, or whether protein binding changes DNA shape. While X-ray structures of protein–DNA complexes are numerous, the structure of naked cognate DNA is seldom available experimentally. We present here an experimental and computational analysis pipeline that uses hydroxyl radical cleavage to map, at single-nucleotide resolution, DNA minor groove width, a recognition feature widely exploited by proteins. For 11 protein–DNA complexes, we compared experimental maps of naked DNA minor groove width with minor groove width measured from X-ray co-crystal structures. Seven sites had similar minor groove widths as naked DNA and when bound to protein. For four sites, part of the DNA in the complex had the same structure as naked DNA, and part changed structure upon protein binding. We compared the experimental map with minor groove patterns of DNA predicted by two computational approaches, DNAshape and ORChID2, and found good but not perfect concordance with both. This experimental approach will be useful in mapping structures of DNA sequences for which high-resolution structural data are unavailable. This approach allows probing of protein family-dependent readout mechanisms.
Rights© The Author(s) 2018. Published by Oxford University Press on behalf of Nucleic Acids Research. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
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