Quantitative visualization of DNA G-quadruplex structures in human cells

• Published in: Nature chemistry Vol. 5; no. 3; pp. 182 - 186
• Main Authors: Biffi, Giulia, Tannahill, David, McCafferty, John, Balasubramanian, Shankar
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 01.03.2013; Nature Publishing Group
• Subjects: 639/638/45/535; 639/638/92/610; 639/638/92/613; Analytical Chemistry; Animals; Antibodies; Biochemistry; Bone Neoplasms - genetics; Cell Cycle; Cell Line, Tumor; Chemistry; Chemistry/Food Science; Deoxyribonucleic acid; DNA; DNA - chemistry; Enzyme-Linked Immunosorbent Assay; G-Quadruplexes; Genomes; Humans; Inorganic Chemistry; Male; Mammals; Nucleic acids; Organic Chemistry; Osteosarcoma - genetics; Physical Chemistry; Physiology; Proteins; Salmon; Spermatozoa - chemistry; Telomerase; Thermodynamics; Visualization; United Kingdom > UK
• ISSN: 1755-4330; 1755-4349; 1755-4349
• DOI: 10.1038/nchem.1548

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. A structure-specific antibody generated and employed to visualize DNA G-quadruplex structures in human cells shows that these structures are modulated during the cell cycle and can be stabilized by a small-molecule ligand. This provides substantive evidence for endogenous DNA G-quadruplex formation in mammalian cells.