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Quantitative visualization of DNA G-quadruplex structures in human cells

Abstract

Four-stranded G-quadruplex nucleic acid structures are of great interest as their high thermodynamic stability under near-physiological conditions suggests that they could form in cells. Here we report the generation and application of an engineered, structure-specific antibody employed to quantitatively visualize DNA G-quadruplex structures in human cells. We show explicitly that G-quadruplex formation in DNA is modulated during cell-cycle progression and that endogenous G-quadruplex DNA structures can be stabilized by a small-molecule ligand. Together these findings provide substantive evidence for the formation of G-quadruplex structures in the genome of mammalian cells and corroborate the application of stabilizing ligands in a cellular context to target G-quadruplexes and intervene with their function.

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Figure 1: Structure specificity of the BG4 antibody for G-quadruplex structures.
Figure 2: Visualization of DNA G-quadruplex structures in nuclei of human cancer cells.
Figure 3: Localization of G-quadruplex structures in chromosomes.
Figure 4: Modulation of G-quadruplex structures during cell-cycle progression.
Figure 5: Stabilization of endogenous G-quadruplex structures by a small-molecule ligand.

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Acknowledgements

We thank T. Pope for his kind help with the phage display, W. Reik, R. Rodriguez and D. Sanders for stimulating discussions and Cancer Research UK for funding.

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Contributions

G.B. carried out the experiments, J.M. advised on the phage display and G.B., D.T. and S.B. designed the experiments. G.B., D.T. and S.B. co-wrote the manuscript.

Corresponding author

Correspondence to Shankar Balasubramanian.

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The authors declare no competing financial interests.

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Biffi, G., Tannahill, D., McCafferty, J. et al. Quantitative visualization of DNA G-quadruplex structures in human cells. Nature Chem 5, 182–186 (2013). https://doi.org/10.1038/nchem.1548

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