Conformational variability of recombination R-triplex formed by the mammalian telomeric sequence

Abstract

Alignment of three nucleic acids strands, in which the third strand is identical to one of the DNA duplex strands, occurs in various cellular systems. In the case of telomeric t-loops, recognition between the DNA duplex and the homologous single strand is likely to be mediated by proteins through formation of the transient recombination-type R-triplex. Earlier, using 2-aminopurine as a fluorescent reporting base, we evaluated the thermodynamic characteristics of intramolecular R-triplex formed by a mixed nucleotide sequence. Here, we used this approach to explore a propensity of the telomeric TTAGGG repeat to form the R-triplex. The circular dichroism spectral changes detected upon formation of the R-triplex suggest that this process is accompanied by specific conformational changes in DNA, including a local destabilization of the target duplex next to a GGG run revealed by the fluorescence of the reporting 2-aminopurine base. Surprisingly, stability of the R-triplex formed by telomeric sequence depends strikingly on the counter ion, being higher for Na⁺ than for Li⁺. Taken together these findings indicate a significant conformational variability of telomeric DNA in the context of recombination-type R-triplex, a phenomenon of possible biological relevance.

Keywords:

• R-triplex
• telomeric DNA
• thermodynamic stability
• fluorescence
• circular dichroism

Acknowledgements

The authors appreciate helpful discussions with V. L. Florent’ev and M.A. Livshits. The study was supported by Molecular and Cellular Biology Program of the Russian Academy of Sciences. Funding to pay the Open Access publication charges for the article was provided by the National Institutes of Health (USA).