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Nucleic Acids Research, 1995, Vol. 23, No. 10 1722-1728
© 1995

CHEMISTRY

Spectroscopic studies of chimeric DNA-RNA and RNA 29-base intramolecular triple helices

J. Liquier, E. Taillandier, R. Klinck1, E. Guittet1, C. Gouyette2 and T. Huynh-Dinh2,*

Laboratoire CSSB URA CNRS 1430, UFR de Médecine, Université Paris Nord, 74 rue Marcel Cachin, 93012 Bobigny Cedex, France 1lnstitut de Chimie des Substances Naturelles CNRS, 98198 Gif sur Yvette, France 2Unité de Chimie Organique URA CNRS 487, Insitut Pasteur, 28 rue du Docteur Roux, 75724 Paris Cedex 15, France

* To whom correspondence should be addressed

Received February 8, 1995. Revised April 6, 1995. Accepted April 6, 1995.

Fourier transform infrared (FTIR), UV absorption and exchangeable proton NMR spectroscopies have been used to study the formation and stability of two intramolecular phi-dependent triple helices composed by a chimeric 29mer DNA-RNA (DNA double strand and RNA third strand) or by the analogous 29mer RNA. In both cases decrease of pH induces formation of a triple helical structure containing either rU*dA.dT and rC+*dG.dC or rU*rA.rU and rC+*rG.rC triplets. FTIR spectroscopy shows that exclusively N-type sugars are present in the triple helix formed by the 29mer RNA while both N- and S-type sugars are detected in the case of the chimeric 29mer DNA-RNA triple helix. Triple helix formation with the third strand RNA and the duplex as DNA appears to be associated with the conversion of the duplex part from a B-form secondary structure to one which contains partly A-form sugars. Thermal denaturatlon experiments followed by UV spectroscopy show that a major stabilization occurs upon formation of the triple helices. Monophasic melting curves indicate a simultaneous disruption of the Hoogsteen and Watson-Crick hydrogen bonds in the intramolecular triplexes when the temperature is increased. This is in agreement with imino proton NMR spectra recorded as a function of temperature. Comparison with experiments concerning intermolecular triplexes of Identical base and sugar composition shows the Important role played by the two tetrameric loops In the stabilization of the intramolecular triple helices studied.


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