Kinetics of DNA hydration

The hydration of the d(CGCGAATTCGCG) B-DNA duplex in solution was studied by nuclear magnetic relaxation dispersion (NMRD) of the water nuclei H-1, H-2, and O-17, and by nuclear Overhauser effects (NOEs) in high-resolution two-dimensional H-1 NMR spectra. By comparing results from the free duplex with those from its complex with netropsin, water molecules in the ''spine of hydration'' in the AATT region of the minor groove could be distinguished from hydration water elsewhere In the duplex. The H-2 and O-17 relaxation dispersions yield a model-independent residence time of 0.9(+/-0.1) ns at 4 degrees C for five highly ordered water molecules in the spine. When corrected for frequency offset effects, the NOE data yield the same residence time as the NMRD data, giving credence to both methods. At 27 degrees C, the residence time is estimated to 0.2 ns, a factor of 40 shorter than the tumbling time of the duplex. The NMRD data show that all water molecules associated with the duplex, except the fi iie molecules in the spine, have residence times significantly shorter than 1 ns at 4 degrees C. There is thus no long-lived hydration structure associated. with the phosphate backbone. In contrast to H-2 and O-17, the H-1 relaxation dispersion is dominated by labile DNA pro-tons and therefore provides Little information about DNA hydration. (C) 1997 Academic Press Limited.