DNA conformational change in Gal repressor-operator complex: involvement of central G-C base pair(s) of dyad symmetry

Abstract

Gal repressor dimer binds to two gal operator sites, OE and OI, which are 16 bp long similar sequences with hyphenated dyad symmetries (11,12). Repressor occupation hinders the reactivity of the N7 atoms in the major groups of guanines, located at positions 1, 3, and 8, and the rotational 1'', 3'' and 8'' of the symmetries. We have shown that Gal repressor binding to OE ior OI DNA fragments increases the circular dichroism (CD) spectral peak in the 270 to 300 nm range. The CD change is similar to that observed for Lac repressor binding to its operator site (14). It is consistent with a DNA conformational change during complex formation between Gal repressor and OE and OI DNA. The CD spectral change was not observed when the central 8,8'' G.sbd.C base pairs in the DNA-protein complex were replaced by A-T base pairs, whereas substitution of the 1,1'' G.sbd.C base pairs do show the accompanying increase in the spectra during repressor binding. The absence of CD change of the Gal repressor complex with DNA mutated at the 8,8'' base pairs suggest that the central G.sbd.C base pairs are required for the repressor induced conformational change.