H-1- and P-31-NMR and UV-absorption studies were carried out with the
oligonucleotide strands d(AGCT-TATC-ATC-GATAAGCT) (-ATC-) and d(AGCTTA
TC-GAT-GATAAGCT) (-GAT-) contained in the strongest and salt resistant
cleavage site for topoisomerase II in pBR322 DNA. We found that the t
wo oligonucleotides were stabilized under a hairpin structure characte
rized by a eight base pair stem and a three base loop at low DNA and s
alt concentrations. In such experimental conditions, only the -GAT- ol
igonucleotide displayed a partial homoduplex structure in slow equilib
rium with its folded structure. Temperature dependencies of imino prot
ons showed that the partial homoduplex of -GAT- melted at a lower temp
erature than the hairpin structure. It was suggested that the appearan
ce of the partial homoduplex in -GAT- is related to the formation of t
wo stabilizing (G-T) mismatched base pairs in the central loop of this
structure. Finally, it was inferred from the dispersion of chemical s
hifts in the P-31-NMR spectra that the distortions affecting the backb
one of the hairpin loop are larger in the case of -ATC-compared with -
GAT-. At the same time NOEs proved that the base stacking was stronger
within the loop of the -ATC-hairpin.