Binding of oligonucleotides to a viral hairpin forming RNA triplexes with parallel G*G·C triplets

Abstract

Infrared and UV spectroscopies have been used to study the assembly of a hairpin nucleotide sequence (nucleotides 3-30) of the 5′ non-coding region of the hepatitis C virus RNA (5′-GGCGGGGAUUAUC-CCCGCUGUGAGGCGG-3′) with a RNA 20mer ligand (5′-CCGCCUCACAAAGGUGGGGU-3′) in the presence of magnesium ion and spermidine. The resulting complex involves two helical structural domains: the first one is an intermolecular duplex stem at the bottom of the target hairpin and the second one is a parallel triplex generated by the intramolecular hairpin duplex and the ligand. Infrared spectroscopy shows that N-type sugars are exclusively present in the complex. This is the first case of formation of a RNA parallel triplex with purine motif and shows that this type of targeting RNA strands to viral RNA duplexes can be used as an alternative to antisense oligonucleotides or ribozymes.