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Nucleic Acids Research, 1990, Vol. 18, No. 18 5387-5391
© 1990

CHEMISTRY

Long (dA)n.(dT)n tracts can form intramolecular triplexes under superhelical stress

Keith R. Fox

Department of Physiology and Pharmacology, University of Southampton Bassett Crescent East, Southampton SO9 3TU, UK

Received July 9, 1990. Revised August 24, 1990. Accepted August 24, 1990.

Plasmids containing long tracts of (dA)n.(dT)n have been prepared and their conformations examined in linear and supercoiled DNA using a series of chemical and enzymic probes which are known to be sensitive to unusual DNA structures. Under superhelical stress and in the presence of magnesium the sequence T69.A69 adopts a conformation at pH 8.0 consistent with the formation of an intramolecular DNA triplex. Site specific cleavage of the supercoiled plasmid by single-strand specific nucleases occurs within the A.T insert; the 5'-end of the purine strand is sensitive to reaction with diethylpyrocarbonate while the central 5–6 bases of the pyrimidine strand are reactive to osmium tetroxide. By contrast shorter inserts of A33.T33 and A23.T23 do not appear to form unusual structures.


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