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Published online 11 May 2004

Nucleic Acids Research, 2004, Vol. 32, No. 8 2598-2606
© 2004 Oxford University Press

Influence of loop size on the stability of intramolecular DNA quadruplexes

Antonina Risitano and Keith R. Fox*

School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK

*To whom correspondence should be addressed. Tel: +44 23 8059 4374; Fax: +44 23 8059 4459; Email: K.R.Fox{at}soton.ac.uk
Present address: Antonina Risitano, School of Medicine, New York University, 550 First Avenue, New York, NY 10016, USA

Received February 11, 2004; Revised and Accepted April 19, 2004

We have determined the stability of intramolecular DNA quadruplexes in which the four G3-tracts are connected by non-nucleosidic linkers containing propanediol, octanediol or hexaethylene glycol, replacing the TTA loops in the human telomeric repeat sequence. We find that these sequences all fold to form intramolecular complexes, which are stabilized by lithium < sodium < potassium. Quadruplex stability increases in the order propanediol < hexaethylene glycol < octanediol. The shallower shape of the melting profile with propanediol linkers and its lower dependency on potassium concentration suggests that this complex contains fewer stacks of G-quartets. The sequence with octanediol linkers displays a biphasic melting profile, suggesting that it can adopt more than one stable structure. All these complexes display melting temperatures above 310 K in the presence of 10 mM lithium, without added potassium, in contrast to the telomeric repeat sequence. These complexes also fold much faster than the telomeric repeat and there is little or no hysteresis between their melting and annealing profiles. In contrast, the human telomeric repeat sequence and a complex containing two hexaethylene glycol groups in each loop, are less stable and fold more slowly. The melting and annealing profiles for the latter sequence show significant differences, even when heated at 0.2°C min–1. CD spectra for the oligonucleotides containing non-nucleosidic linkers show positive maxima at 264 nm, with negative minima ~244 nm, which are characteristic of parallel quadruplex structures. These results show that the structure and stability of intramolecular quadruplexes is profoundly influenced by the length and composition of the loops.


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