Zn²⁺ selectively stabilizes FdU-substituted DNA through a unique major groove binding motif
2011
Ghosh, Supratim | Salsbury, Freddie R. Jr | Horita, David A. | Gmeiner, William H.
We report, based on semi-empirical calculations, that Zn²⁺ binds duplex DNA containing consecutive FdU-dA base pairs in the major groove with distorted trigonal bipyramidal geometry. In this previously uncharacterized binding motif, O4 and F5 on consecutive FdU are axial ligands while three water molecules complete the coordination sphere. NMR spectroscopy confirmed Zn²⁺ complexation occurred with maintenance of base pairing while a slight hypsochromic shift in circular dichroism (CD) spectra indicated moderate structural distortion relative to B-form DNA. Zn²⁺ complexation inhibited ethidium bromide (EtBr) intercalation and stabilized FdU-substituted duplex DNA (ΔTm > 15°C). Mg²⁺ neither inhibited EtBr complexation nor had as strong of a stabilizing effect. DNA sequences that did not contain consecutive FdU were not stabilized by Zn²⁺. A lipofectamine preparation of the Zn²⁺-DNA complex displayed enhanced cytotoxicity toward prostate cancer cells relative to the individual components prepared as lipofectamine complexes indicating the potential utility of Zn²⁺-DNA complexes for cancer treatment.
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