King, GA;
Biebricher, AS;
Heller, I;
Peterman, EJG;
Wuite, GJL;
(2018)
Quantifying Local Molecular Tension Using Intercalated DNA Fluorescence.
Nano Letters
, 18
(4)
pp. 2274-2281.
10.1021/acs.nanolett.7b04842.
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Abstract
The ability to measure mechanics and forces in biological nanostructures, such as DNA, proteins and cells, is of great importance as a means to analyze biomolecular systems. However, current force detection methods often require specialized instrumentation. Here, we present a novel and versatile method to quantify tension in molecular systems locally and in real time, using intercalated DNA fluorescence. This approach can report forces over a range of at least ∼0.5−65 pN with a resolution of 1−3 pN, using commercially available intercalating dyes and a general-purpose fluorescence microscope. We demonstrate that the method can be easily implemented to report double-stranded (ds)DNA tension in any single-molecule assay that is compatible with fluorescence microscopy. This is particularly useful for multiplexed techniques, where measuring applied force in parallel is technically challenging. Moreover, tension measurements based on local dye binding offer the unique opportunity to determine how an applied force is distributed locally within biomolecular structures. Exploiting this, we apply our method to quantify the positiondependent force profile along the length of flow-stretched DNA and reveal that stretched and entwined DNA molecules mimicking catenated DNA structures in vivodisplay transient DNA−DNA interactions. The method reported here has obvious and broad applications for the study of DNA and DNA−protein interactions. Additionally, we propose that it could be employed to measure forces in any system to which dsDNA can be tethered, for applications including protein unfolding, chromosome mechanics, cell motility, and DNA nanomachines.
| Type: | Article |
|---|---|
| Title: | Quantifying Local Molecular Tension Using Intercalated DNA Fluorescence |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1021/acs.nanolett.7b04842 |
| Publisher version: | https://doi.org/10.1021/acs.nanolett.7b04842 |
| Language: | English |
| Additional information: | This is an open access article published under a Creative Commons Non-Commercial No Derivative Works (CC-BY-NC-ND) Attribution License, which permits copying and redistribution of the article, and creation of adaptations, all for non-commercial purposes https://creativecommons.org/licenses/by-nc-nd/4.0/ |
| Keywords: | force sensor, molecular tension, DNA, intercalators, fluorescence microscopy |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery-pp.ucl.ac.uk/id/eprint/10107760 |
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