Oligonucleotide-Minor Groove Binder 1:2 Conjugates: Side by Side Parallel Minor Groove Binder Motif in Stabilization of DNA Duplex

• Published in: Nucleosides, nucleotides & nucleic acids nucleotides & nucleic acids, 2004-10, Vol.23 (6-7), p.953-968
• Main Authors: Ryabinin, Vladimir A., Boutorine, Alexandre S., Hélène, Claude, Denisov, Alexey Yu, Pyshnyi, Dmitrii V., Sinyakov, Alexandre N.
• Format: Article
• Language: English
• Subjects: Chromatography, High Pressure Liquid; Conjugates; DNA - chemistry; DNA - metabolism; Duplex stability; Electrophoresis, Polyacrylamide Gel; Mass Spectrometry; Minor groove binders; Nuclear Magnetic Resonance, Biomolecular; Nucleic Acid Denaturation; Oligonucleotides; Oligonucleotides - chemistry; Oligonucleotides - metabolism; Sequence specificity; Thermal denaturation
• ISSN: 1525-7770; 1532-2335
• DOI: 10.1081/NCN-200026046

Synthetic polycarboxamides consisting of N-methylpyrrole (Py), N-methylimidazole (Im), N-methyl-3-hydroxypyrrole (Hp) and β-alanine (β) show strong and sequence-specific interaction with the DNA minor groove when they form hairpin structures with side-by-side antiparallel motifs. In the present paper, new conjugates containing two ligands linked to the same terminal phosphate of DNA strand were constructed. The paper describes optimized synthesis and properties of oligonucleotide-linked polyamide strands that insert into the minor groove of a duplex in a parallel or antiparallel orientation. Strong stabilization of DNA duplexes by two attached minor groove ligands is demonstrated by the thermal denaturation method. The unmodified duplex 5′-CGTTTATTp-3′/5′-AATAAACG-3′ melts at 20°C. When one tetra(Py) residue was attached to the first strand of this duplex, denaturation temperature was increased to 46°C; attachment of the second tetra(Py) in a parallel orientation resulted in denaturation temperature of 60°C. It is even higher than in case of "classic" octapyrrole hairpin ligand (Tm = 58°C). Sequence-specific character of stabilization by two conjugated ligands was demonstrated for G:C-containing oligonucleotides attached to tetracarboxamide and octacarboxamide ligands constructed from Py, Im and β units according to established recognition rules (ΔTm = 20°C). The two-strand parallel minor groove binder constructions attached to addressing oligonucleotides could be considered as site-specific ligands recognizing single- and double-stranded DNA similarly to already described hairpin MGB structures with antiparallel orientation of carboxamide units.