Di Antonio, M, Ponjavic, A orcid.org/0000-0002-7561-1127, Radzevičius, A et al. (8 more authors) (2020) Single-molecule visualization of DNA G-quadruplex formation in live cells. Nature Chemistry, 12 (9). pp. 832-837. ISSN 1755-4330
Abstract
Substantial evidence now exists to support that formation of DNA G-quadruplexes (G4s) is coupled to altered gene expression. However, approaches that allow us to probe G4s in living cells without perturbing their folding dynamics are required to understand their biological roles in greater detail. Herein, we report a G4-specific fluorescent probe (SiR-PyPDS) that enables single-molecule and real-time detection of individual G4 structures in living cells. Live-cell single-molecule fluorescence imaging of G4s was carried out under conditions that use low concentrations of SiR-PyPDS (20 nM) to provide informative measurements representative of the population of G4s in living cells, without globally perturbing G4 formation and dynamics. Single-molecule fluorescence imaging and time-dependent chemical trapping of unfolded G4s in living cells reveal that G4s fluctuate between folded and unfolded states. We also demonstrate that G4 formation in live cells is cell-cycle-dependent and disrupted by chemical inhibition of transcription and replication. Our observations provide robust evidence in support of dynamic G4 formation in living cells.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author(s), under exclusive licence to Springer Nature Limited 2020. This is an author produced version of an article published in Nature Chemistry. Uploaded in accordance with the publisher's self-archiving policy. |
Keywords: | DNA; Nucleic acids |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Physics and Astronomy (Leeds) > Molecular & Nanoscale Physics |
Depositing User: | Symplectic Publications |
Date Deposited: | 03 Aug 2020 14:59 |
Last Modified: | 20 Jan 2021 01:39 |
Status: | Published |
Publisher: | Nature Research |
Identification Number: | 10.1038/s41557-020-0506-4 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:163757 |