Identification of Three-Way DNA Junction Ligands through Screening of Chemical Libraries and Validation by Complementary in Vitro Assays.
Katerina DuskovaJérémy LamarcheSouheila AmorCoralie CaronNicolas QueyriauxMarie GaschardMarie-José PenouilhGuillaume de RobillardDominique DelmasCharles H DevillersAnton GranzhanMarie-Paule Teulade-FichouMurielle Chavarot-KerlidouBruno TherrienSébastien BrittonDavid MonchaudPublished in: Journal of medicinal chemistry (2019)
The human genome is replete with repetitive DNA sequences that can fold into thermodynamically stable secondary structures such as hairpins and quadruplexes. Cellular enzymes exist to cope with these structures whose stable accumulation would result in DNA damage through interference with DNA transactions such as transcription and replication. Therefore, the chemical stabilization of secondary DNA structures offers an attractive way to foster DNA transaction-associated damages to trigger cell death in proliferating cancer cells. While much emphasis has been recently given to DNA quadruplexes, we focused here on three-way DNA junctions (TWJ) and report on a strategy to identify TWJ-targeting agents through a combination of in vitro techniques (TWJ-screen, polyacrylamide gel electrophoresis, fluorescence resonance energy transfer-melting, electrospray ionization mass spectrometry, dialysis equilibrium, and sulforhodamine B assays). We designed a complete workflow and screened 1200 compounds to identify promising TWJ ligands selected on stringent criteria in terms of TWJ-folding ability, affinity, and selectivity.
Keyphrases
- single molecule
- circulating tumor
- cell free
- energy transfer
- high resolution
- cell death
- dna damage
- mass spectrometry
- high throughput
- endothelial cells
- chronic kidney disease
- transcription factor
- genome wide
- high frequency
- drug delivery
- signaling pathway
- liquid chromatography
- molecular dynamics simulations
- capillary electrophoresis
- molecular dynamics
- hyaluronic acid
- induced pluripotent stem cells
- cancer therapy