Intrusion of Counterions into the Spine of Hydration in the Minor Groove of B-DNA: Fractional Occupancy of Electronegative Pockets

A sequence of ordered solvent peaks in the electron density map of the minor groove region of ApT-rich tracts of the double helix is a characteristic of B-form DNA well established from crystallography. This feature, termed the “spine of hydration”, has been discussed as a stabilizing feature of B-DNA, the structure of which is known to be sensitive to environmental effects. Nanosecond-range molecular dynamics simulations on the DNA duplex of sequence d(CGCGAATTCGCG) have been carried out, including explicit consideration of ∼4000 water molecules and 22 Na+ counterions, and based on the new AMBER 4.1 force field with the particle mesh Ewald summation used in the treatment of long-range interactions. The calculations support a dynamical model of B-DNA closer to the B form than any previously reported. Analysis of the dynamical structure of the solvent revealed that, in over half of the trajectory, a Na+ ion is found in the minor groove localized at the ApT step. This position, termed herein the “ApT pocket”, was noted previously (Lavery, R.; Pullman, B. J. Biomol. Struct. Dyn. 1985, 5, 1021) to be of uniquely low negative electrostatic potential relative to other positions of the groove, a result supported by the location of a Na+ ion in the crystal structure of the dApU miniduplex [Seeman, N.; et al. J. Mol. Biol. 1976, 104, 109) and by additional calculations described herein based on continuum electrostatics. The Na+ ion in the ApT pocket interacts favorably with the thymine O2 atom on opposite strands of the duplex and is well articulated with the water molecules which constitute the remainder of the minor groove spine. This result indicates that counterions may intrude on the minor groove spine of hydration on B-form DNA and subsequently influence the environmental structure and thermodynamics in a sequence-dependent manner. The observed narrowing of the minor groove in the AATT region of the d(CGCGAATTCGCG) structure may be due to direct binding effects and also to indirect modulation of the electrostatic repulsions that occur when a counterion resides in the minor groove “AT pocket”. The idea of localized complexation of otherwise mobile counterions in electronegative pockets in the grooves of DNA helices introduces a heretofore mostly unappreciated source of sequence-dependent effects on local conformational, helicoidal, and morphological structure and may have important implications in understanding the functional energetics and specificity of the interactions of DNA and RNA with regulatory proteins, pharmaceutical agents, and other ligands.