Improving the accuracy of NMR structures of DNA by means of a database potential of mean force describing base-base positional interactions

NMR structure determination of nucleic acids presents an intrinsically diff icult problem since the density of short interproton distance contacts is r elatively low and limited to adjacent base pairs. Although residual dipolar couplings provide orientational information that is clearly helpful, they do not provide translational information of either a short-range (with the exception of proton-proton dipolar couplings) or long-range nature. As a co nsequence, the description of the nonbonded contacts has a major impact on the structures of nucleic acids generated from NMR data. In this paper, we describe the derivation of a potential of mean force derived from all high- resolution (2 Angstrom or better) DNA crystal structures available in the N ucleic Acid Database (NDB) as of May 2000 that provides a statistical descr iption, in simple geometric terms, of the relative positions of pairs of ne ighboring bases (both intra- and interstrand) in Cartesian space. The purpo se of this pseudopotential, which we term a DELPHIC base-base positioning p otential, is to bias sampling during simulated annealing refinement to phys ically reasonable regions of conformational space within the range of possi bilities that are consistent with the experimental NMR restraints. We illus trate the application of the DELPHIC base-base positioning potential to the structure refinement of a DNA dodecamer, d(CGCCAATTCGCG)(2), for which NOE and dipolar coupling data have been measured in solution and for which cry stal structures have been determined. We demonstrate by cross-validation ag ainst independent NMR observables (that is, both residual dipolar couplings and NOE-derived intereproton distance restraints) that the DELPHIC base-ba se positioning potential results in a significant increase in accuracy and obviates artifactual distortions in the structures arising from the Limitat ions of conventional descriptions of the nonbonded contacts in terms of eit her Lennard-Jones van der Waals and electrostatic potentials or a simple va n der Waals repulsion potential. We also demonstrate, using experimental NM R data for a complex of the male sex determining factor SRY with a duplex D NA 14mer, which includes a region of highly unusual and distorted DNA, that the DELPHIC base-base positioning potential does not in any way hinder unu sual interactions and conformations from being satisfactorily sampled and r eproduced. We expect that the methodology described in this paper for DNA c an be equally applied to RNA, as well as side chain-side chain interactions in proteins and protein-protein complexes, and side chain-nucleic acid int eractions in protein-nucleic acid complexes. Further, this approach should be useful not only for NMR structure determination but also for refinement of low-resolution (3-3.5 Angstrom) X-ray data.