Issue 6, 2015

G-quadruplex DNA and ligand interaction in living cells using NMR spectroscopy

Abstract

Gathering structural information from biologically relevant molecules inside living cells has always been a challenging task. In this work, we have used multidimensional NMR spectroscopy to probe DNA G-quadruplexes inside living Xenopus laevis oocytes. Some of these structures can be found in key regions of chromosomes. G-quadruplexes are considered potential anticancer therapeutic targets and several lines of evidence indirectly point out roles in key biological processes, such as cell proliferation, genomic instability or replication initiation. However, direct demonstrations of the existence of G-quadruplexes in vivo are scarce. Using SOFAST-HMQC type spectra, we probed a tetramolecular G-quadruplex model made of d(TG4T)4 inside living Xenopus laevis oocytes. Our observations lead us to conclude that the quadruplex structure is formed within the cell and that the intracellular environment preferentially selects a conformation that most resembles the one found in vitro under KCl conditions. We also show for the first time that specific ligands targeting G-quadruplexes can be studied using high resolution NMR directly inside living cells, opening new avenues to study ligand binding discrimination under physiologically relevant conditions with atomic detail.

Graphical abstract: G-quadruplex DNA and ligand interaction in living cells using NMR spectroscopy

Supplementary files

Article information

Article type
Edge Article
Submitted
12 Dec 2014
Accepted
13 Jan 2015
First published
14 Jan 2015
This article is Open Access

All publication charges for this article have been paid for by the Royal Society of Chemistry
Creative Commons BY license

Chem. Sci., 2015,6, 3314-3320

Author version available

G-quadruplex DNA and ligand interaction in living cells using NMR spectroscopy

G. F. Salgado, C. Cazenave, A. Kerkour and J. Mergny, Chem. Sci., 2015, 6, 3314 DOI: 10.1039/C4SC03853C

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence. You can use material from this article in other publications without requesting further permissions from the RSC, provided that the correct acknowledgement is given.

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