Impact of Intercalators on the Properties of DNA Analyzed by Molecular Dynamics Simulations

Abstract Intercalation of small molecules between DNA base pairs affects DNA conformation, disrupting essential cellular processes, including replication, transcription, and repair. We investigated conformational changes in 18-mer DNA upon intercalation of doxorubicin, SYBR Gold, and YOYO-1 using extensive molecular dynamics simulations. Two main patterns for the intercalation were identified: RISE-type intercalation occurs between adjacent base pairs and extends the DNA helix with decreased twist angles, whereas BPEV-type intercalation proceeds through base-pair eversion without significant DNA extension. Kinetic analysis revealed that association rates for intercalation followed the order: first YO moiety (monointercalation) > SYBR Gold > doxorubicin > YOYO-1 (bis-intercalation). The free energy landscape showed that forces at DNA termini reached up to 117 pN during stretching. Notably, base pairs adjacent to intercalators were protected from strand separation, accompanied by additional helical unwinding. MM-GBSA/PBSA analysis revealed that the driving force for intercalation is the stacking energy, and the binding affinity was highest for minor-groove binding. Persistence length decreased with single-molecule binding but recovered with two molecules due to their electrostatic repulsion. Mechanical properties of intercalated DNA showed position dependence, demonstrating that multiple intercalation modes coexist in solution. The heterogeneous nature of intercalation explains why experimental measurements reflect ensemble averages, rather than single binding configurations.