Evidence for (PNA)2/DNA triplex structure upon binding of PNA to dsDNA by strand displacement

The binding of PNA (peptide nucleic acid) T₂CT₂CT₄-LysNH₂ to the double-stranded DNA target 5′ -A₂GA₂GA₄ was studied by KMnO₄ and dimethylsulfate (DMS) probing. It is found that upon sequence-specific strand displacement binding of the PNA to the dsDNA target concomitant protection of the N-7 of guanines within the target takes place. It is furthermore shown that the binding of this PNA is more efficient at pH 5.5 that at pH 6.5 and very inefficient at pH 7.5. These results clearly indicate that C⁺G Hoogsteen base pairing is present and important for binding and that the strand displacement complex therefore involves a PNA·DNA-PNA triplex.