title: Non-covalent duplex to duplex crosslinking of DNA in solution revealed by single molecule force spectroscopy.
abstract: Small molecules that interact with DNA, disrupting the binding of transcription factors or crosslinking DNA into larger structures, have significant potential as cancer therapies and in nanotechnology. Bisintercalators, including natural products such as echinomycin and rationally designed molecules such as the bis-9-aminoacridine-4-carboxamides, are key examples. There is little knowledge of the propensity of these molecules to crosslink duplex DNA. Here we use single molecule force spectroscopy to assay the crosslinking capabilities of bisintercalators. We show that bis-9-aminoacridine-4-carboxamides with both rigid and flexible linkers are able to crosslink duplex strands of DNA, and estimate the equilibrium free energy of a 9-aminoacridine-4-carboxamide bisintercalator from DNA at 5.03 kJ mol(-1). Unexpectedly, we find that echinomycin and its synthetic analogue TANDEM are capable of sequence-specific crosslinking of the terminal base pairs of two duplex DNA strands. In the crowded environment of the nucleosome, small molecules that crosslink neighbouring DNA strands may be expected to have significant effects on transcription, while a small molecule that facilitates sequence-specific blunt-end ligation of DNA may find applications in the developing field of DNA nanotechnology.
authors.affiliation: School of Pharmacy, University of East Anglia, Norwich Research Park, Norwich NR4 7TJ, UK. m.searcey@uea.ac.uk a.round@uea.ac.uk.
authors.forename: Benjamin D
authors.initials: BD
authors.lastname: Rackham
authors.forename: Lesley A
authors.initials: LA
authors.lastname: Howell
authors.forename: Andrew N
authors.initials: AN
authors.lastname: Round
authors.forename: Mark
authors.initials: M
authors.lastname: Searcey
mesh_terms: D000609:Aminoacridines / Q000737:chemistry* / Q000494:pharmacology; D001483:Base Sequence; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000494:pharmacology; D004247:DNA / Q000737:chemistry*; D004448:Echinomycin / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D007364:Intercalating Agents / Q000737:chemistry* / Q000494:pharmacology; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D019384:Nucleic Acid Synthesis Inhibitors / Q000737:chemistry / Q000494:pharmacology; D013057:Spectrum Analysis
chemical_list: D000609:Aminoacridines; D003432:Cross-Linking Reagents; D007364:Intercalating Agents; D019384:Nucleic Acid Synthesis Inhibitors; C054505:9-aminoacridine-4-carboxamide; D004448:Echinomycin; D004247:DNA