ENERGETICS OF BASE-PAIR OPENING IN A DNA DODECAMER CONTAINING AN A(3)T(3) TRACT

Nuclear magnetic resonance spectroscopy has been used to characterize the kinetics and energetics of opening of base pairs in the DNA dodecamer [d(CGCAAATTTGCG)](2). The dodecamer contains an A(3)T(3) tract that induces intrinsic curvature of the helix axis. previous studies from this and other laboratories have shown that the kinetics of base pair opening in A(3)T(3) tracts is unique: the opening rates of the A . T base pairs in the interior of the tract are much lower than that of the A . T base pair at the 5'-end of the tract. In the present work, we have investigated the energetics of the pathways for opening of the A . T base pairs in the A(3)T(3) tract. The energetic parameters of the activated state(s) are obtained from the temperature dependence of the opening rate constants. The lower opening rates for the A . T base pairs situated in the interior of the tract are shown to originate from higher activation enthalpies which are compensated, in part, by increases in the activation entropies. We have also obtained an energetic characterization of the open state(s) of the A . T base pairs in the dodecamer by measuring the equilibrium constants for base pair opening and their temperature dependence. The results suggest that the transitions from closed to open state(s) in the A . T base pairs of the A(3)T(3) tract are energetically similar.