PARALLEL AND ANTIPARALLEL G-DNA STRUCTURES FROM A COMPLEX TELOMERIC SEQUENCE

We investigated the formation in vitro of higher order structures by a DNA oligomer containing the terminal motif TGTG3TGTGTGTG3, derived from the Saccharomyces telomeric consensus, in order to (a) understand why certain cations favor the formation of parallel-stranded (G4 and G8) G-DNA structures, while others favor foldback, antiparallel structures (G'2) and (b) probe the structures of G-DNAs formed by this telomeric sequence, which is more complex than its well-studied counterparts from the protozoans oxytricha and tetrahymena. We find that dramatic switches in the formation of G4 versus G'2 structures occur in solutions of not only the group Ia cations, Li+-Cs+, but also in those of the group IIa cations, Mg2+-Ba2+. These data and the temperature-dependent formation and destruction of the different structures lend support to the kinetic scheme of Sen and Gilbert (1990), by which rapidly forming G'2 structures accumulate in highly stabilizing potassium (and strontium) solutions at the expense of the thermodynamically more stable G4 structures. Both the G4 and the G'2 complexes formed by the Saccharomyces sequence show novel structural features. Protection and interference experiments with dimethyl sulfate and potassium permanganate reveal that the core of alternating thymines and guanines within the telomeric motif plays a critical role in the stabilization of the parallel G4 structure, but not of the antiparallel G'2. Very likely, in the G4 complex, this GT core forms a novel higher order arrangement of alternating G and T quartets, the latter possibly comparable to the U quartets described by Cheong and Moore (1992) in their NMR study of the higher order structure formed by rUG4U.