Comparative stability of eight different triple helices formed by differently modified DNA or RNA pyrimidine strands and a DNA hairpin

The comparative stability of eight different triplexes constituted with 16-mer pyrimidine-modified oligodeoxynucleotides (wild-type ODN, PS-ODN, alpha-ODN, or alpha-PS-ODN) or oligoribonucleotides (wild-type ORN, alpha-ORN, 4'-thio-ORN, or 2'-O-MeORN) and a DNA hairpin, termed H36, was studied in five different buffers by UV melting curve analysis, The composition of buffers varied in pH (5.5 and 6.5), in salt concentration (100 mM and 1 M Na+), and in the presence or absence of divalent cation (0 or 3 mM Mg2+) or, Spermine (0 or 1 mM), At pH 5.5, the eight triplexes are formed with T-m values ranging from 24.7 degrees C to 50.9 degrees C (Delta G(298K) between -8.1 and -16.8 kcal/mol). At pH 6.5, the triplexes are less stable, and thus 4'-thio-ORN and PS-ODN showed broad transitions that did not allow us to conclude tripler formation, An increase of salt concentration or the presence of spermine stabilizes the triplexes, whereas Mg2+ has a destabilizing effect (excepted for alpha-ORN), In general ORN:H36 and 2'-O-MeORN:H36 triplexes were the most stable, Finally, introduction of alpha-anomeric nucleosides led to an alpha-ORN analog that showed low binding with H36 and to alpha-ODN and alpha-PS-ODN analogs, Triplexes formed with alpha-ODN were slightly less stable than those formed with unmodified ODN, Surprisingly, introduction of phosphorothioate in an alpha analog led only to a low destabilization.