Indirect DNA Readout by an H-NS Related Protein: Structure of the DNA Complex of the C-Terminal Domain of Ler
Figure 8
The DNA-binding domains of Ler and H-NS share a similar indirect DNA readout mechanism.
(A) EMSA (1.5% agarose) of the −225 to +121 LEE2 fragment (80 ng) with increasing concentrations of wild type and R114G H-NS proteins. (B) DNA titrations of CT-H-NS followed by NMR. Expansions of 1H-15N HSQC spectra of CT-H-NS in the presence of the 10 bp duplexes AATT (top left, 0, 0.5, 1, 2, 3 and 4.5 equivalents) or TATA (top right, 0, 1, 2, 3, 4.5 and 6 equivalents). The DNA-dependent shifts of selected cross-peaks were fitted to a 1∶1 model (bottom), supported by the strict linear displacement of the cross-peaks during the titration. (C) CT-Ler and CT-H-NS binding to the −225 to +121 LEE2 fragment (20 ng) followed by EMSA on a 7% polyacrylamide gel.