Comparative studies of duplex and triplex formation of 2'-5' and 3'-5' linked oligoribonucleotides.

Abstract

We have studied double and triple helix formation between 2'-5' or 3'-5' linked oligoriboadenylates and oligoribouridylates with chain length 7 or 10 by CD spectrometry. The complex formation depends on the type of linkage of oligoribonucleotides, chain length, concentration and molar ratio of the strands, temperature and the cationic concentration. Mixture of any linkage isomers of oligo(rA) and oligo(rU) in 1:1 molar ratio form duplex at 0.1 M NaCl. The duplex stability largely depends on the type of the linkages and is in the following order, [3'-5'] oligo(rA)-[3'-5'] oligo(rU) > [2'-5'] oligo(rA)-[3'-5'] oligo(rU) > [3'-5'] oligo(rA)-[2'-5'] oligo(rU) > [2'-5'] oligo(rA)-[2'-5'] oligo(rU). The higher cationic concentrations, 0.5 M MgCl2, stabilize the complex and either duplex or triplex is formed depending on the input strand ratio and the type of linkage. Thermodynamic parameters, DH and DS, for the complex formation between linkage isomers of oligo(rA) and oligo(rU) showed a linear relationship indicating an enthalpy-entropy compensation phenomena. The duplex and triplex composed of [2'-5'] oligo(rA) and [2'-5'] oligo(rU) exhibit different CD spectra compared to those of any others containing 3'-5' linkage, suggesting that the fully 2'-5' duplex and triplex may possess a unique conformation. We describe prebiological significance of the linkage isomers of RNA and selection of the 3'-5' linkage against 2'-5 linkage.