Comparison of POU/HMG complexes formed on FGF4 and UTF1.
(A, top) Model of POU/HMG/FGF4 (POU domain of Oct1 or Oct4;
left), compared with the model of Oct4-POU/HMG/UTF1
(right). The figure illustrates that different spacing between the
binding sites for the POU and HMG domains within FGF4 and
UTF1 enhancers causes formation of different heterodimeric
interfaces. (Bottom) Close-up views of the HMG/POUS
interfaces on FGF4 (left) and UTF1
(right). The POU/HMG/UTF1 model suggests involvement of
helix 3 instead of the C terminus of the HMG domain to form the
HMG-POUS interface, whereas the same surface patch of the
POUS domain appears to be involved in both interfaces. The DNA
molecules are depicted with a transparent surface and are brown. Coloring of
the FGF4 and UTF1 DNA sequences is according to their POU-
and Sox2-HMG-binding sites. Notice the difference in spacing of these two
sites in the two enhancers. (B) To validate the homology models in
A, mutations in Sox2-HMG were designed to selectively interfere with
ternary complex formation on the FGF4 (m1) or on the UTF1
enhancers (m2 and m3); (m1) R75E; (m2) K57E,R60E; (m3) R60E,M64E; (mut) mutant
version of Oct4 POU (I21Y,D29E). In agreement with both models, m1
specifically impaired heterodimerization on FGF4, whereas m2 and m3
specifically abrogated heterodimerization on UTF1, but Oct4 mut
affected both.