Crowding-induced hybridization of single DNA hairpins

Baltierra-Jasso, L. E., Morten, M. J., Laflor, L., Quinn, S. D. and Magennis, S. W. (2015) Crowding-induced hybridization of single DNA hairpins. Journal of the American Chemical Society, 137(51), pp. 16020-16023. (doi: 10.1021/jacs.5b11829) (PMID:26654490)

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It is clear that a crowded environment influences the structure, dynamics, and interactions of biological molecules, but the complexity of this phenomenon demands the development of new experimental and theoretical approaches. Here we use two complementary single-molecule FRET techniques to show that the kinetics of DNA base pairing and unpairing, which are fundamental to both the biological role of DNA and its technological applications, are strongly modulated by a crowded environment. We directly observed single DNA hairpins, which are excellent model systems for studying hybridization, either freely diffusing in solution or immobilized on a surface under crowding conditions. The hairpins followed two-state folding dynamics with a closing rate increasing by 4-fold and the opening rate decreasing 2-fold, for only modest concentrations of crowder [10% (w/w) polyethylene glycol (PEG)]. These experiments serve both to unambiguously highlight the impact of a crowded environment on a fundamental biological process, DNA base pairing, and to illustrate the benefits of single-molecule approaches to probing the structure and dynamics of complex biomolecular systems.

Item Type:Articles
Glasgow Author(s) Enlighten ID:Quinn, Dr Steven and Magennis, Dr Steven and Morten, Dr Michael
Authors: Baltierra-Jasso, L. E., Morten, M. J., Laflor, L., Quinn, S. D., and Magennis, S. W.
College/School:College of Science and Engineering > School of Chemistry
Journal Name:Journal of the American Chemical Society
Publisher:American Chemical Society
ISSN (Online):1520-5126
Published Online:11 December 2015
Copyright Holders:Copyright © 2015 American Chemical Society
First Published:First published in Journal of the American Chemical Society 137(51):16020-16023
Publisher Policy:Reproduced under a creative commons license

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
647971Speeding and stuttering: analysing the dynamics of DNA replication at the single molecule levelSteven MagennisBiotechnology and Biological Sciences Research Council (BBSRC)BB/K001957/1SCHOOL OF CHEMISTRY
657181From nature to nano: structure, dynamics and reactivity of DNA three-way junctionsSteven MagennisEngineering & Physical Sciences Research Council (EPSRC)EP/L027003/1SCHOOL OF CHEMISTRY