SECONDARY STRUCTURE OF THE DNA-BINDING DOMAIN OF THE C-MYB ONCOPROTEIN IN SOLUTION - A MULTIDIMENSIONAL DOUBLE AND TRIPLE HETERONUCLEAR NMR-STUDY

The DNA-binding domain of the c-Myb oncoprotein contains two repeats, R2 and R3, both of which have been proposed to be related to the helix-turn-helix(HTH) motif. As a first step towards determination of the three-dimensional structure of this domain and of the mode of interaction with the DNA, we have undertaken multidimensional heteronuclear NMR studies using uniformly N-15-labeled and C-13, N-15 double-labeled R2R3 and, a selectively N-15-enriched sample on all lysine, histidine and leucine residues of R2R3. We present almost complete assignments of the backbone H-1, N-15 and C-13alpha atoms and determine the secondary structure of R2R3 in solution. The R3 repeat is composed of three helices (residues 62-75, 78-85 and 91-100) while for the R2 repeat only two helices are found (residues 10-23 and 28-34). The remaining C-terminal part of the R2 repeat, predicted to be helical and part of the HTH motif, undergoes intermediate conformational exchange processes. Stabilization of this segment might occur upon binding to DNA.