Structural basis for DNA bending.
AUTOR(ES)
Nadeau, J G
RESUMO
We report proton NMR studies on DNA oligonucleotides that contain A tracts of lengths known to produce various degrees of bending. Spectra of duplexes in the series 5'-(GGCAnCGG).(CCGTnGCC) (n = 3, 4, 5, 7, 9) reveal substantial structural changes within the An.Tn tract as its length is increased. Chemical-shift comparisons show that A tracts with fewer than about seven members do not contain regions of uniform [or poly(dA).poly(dT)-like] structure. Long An tracts (n greater than or equal to 7) appear to consist of an internal segment of homopolymeric conformation flanked by regions of transitional structure that occupy about four A.T pairs on the 5' side and two A.T pairs perhaps the directly adjacent G.C pair on the 3' side. In shorter duplexes (n less than 7), these two transitional regions overlap and an apparent mutual incompatibility causes length-dependent changes that are most pronounced near the 3' end. Throughout the series, there is a striking monotonic relationship between the location of an A.T pair in the A tract and the relative position of its ThyH3 resonance. The direction of the chemical-shift dispersion is opposite to that expected from consideration of ring-current effects alone; this discrepancy suggests a gradual decrease in ThyH3...N1Ade hydrogen-bond length as one moves from the 5' to the 3' end of the A tract and from short to long A tracts. Nuclear Overhauser effect measurements reveal that the interproton distances AdeH2...H1'Ade and AdeH2...H1'Thy vary along each A tract, except in the central regions of the longer ones where they are fairly constant and in good agreement with the poly(dA).poly(dT) structure proposed by Lipanov, A.A. & Chuprina, V. P. [(1987) Nucleic Acids Res. 15, 5833-5844]. This model features a substantial negative base-pair tilt, which has been suggested previously as the source of A-tract bending. In contrast, the nuclear Overhauser effect distances are inconsistent with at least one known crystallographic A-tract structure [DiGabriele, A. D., Sanderson, M. R. & Steitz, T. A. (1989) Proc. Natl. Acad. Sci. USA 86, 1816-1820], which lacks appreciable base-pair tilt.
