Direct Measurement of Interhelical DNA Repulsion and Attraction by Quantitative Cross-Linking
2022
Hamilton, Ian | Gebala, Magdalena | Herschlag, Daniel | Russell, Rick
To better understand the forces that mediate nucleic acid compaction in biology, we developed the disulfide cross-linking approach xHEED (X-linking of Helices to measure Electrostatic Effects at Distance) to measure the distance-dependent encounter frequency of two DNA helices in solution. Using xHEED, we determined the distance that the electrostatic potential extends from DNA helices, the dependence of this distance on ionic conditions, and the magnitude of repulsion when two helices approach one another. Across all conditions tested, the potential falls to that of the bulk solution within 15 Å of the major groove surface. For separations of ∼30 Å, we measured a repulsion of 1.8 kcal/mol in low monovalent ion concentration (30 mM Na⁺), with higher Na⁺ concentrations ameliorating this repulsion, and 2 M Na⁺ or 100 mM Mg²⁺ eliminating it. Strikingly, we found full screening at very low Co³⁺ concentrations and net attraction at higher concentrations, without the higher-order DNA condensation that typically complicates studies of helical attraction. Our measurements define the relevant distances for electrostatic interactions of nucleic-acid helices in biology and introduce a new method to propel further understanding of how these forces impact biological processes.
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