Effects of chloroquine on the torsional dynamics and rigidities of linear and supercoiled DNAs at low ionic strength

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Abstract
The magnitude and uniformity of the torsion elastic constant (α) of linear and supercoiled pBR322 DNAs are measured in 3 mM Tris as a function of added chloroquine/basepair ratio (chl/bp) by studying the fluorescence polarization anisotropy of intercalated ethidium dye. The time-resolved FPA is measured using a picosecond dye-laser for excitation and time-correlated single-photon counting detction. For both linear and supercoiled DNAs, α remains uniform except at the very highest chl/bp ratio examined. For the linear DNA, α decreases from 5.0 × 10−12 dyne-cm at chl/bp = 0 to about 3.5 × 10−12 dyne-cm at chl/bp = 0.5, and remains at that value up to chl/bp = 5, whereupon it increases back up to its original value. For the supercoiled DNA, α remains constant at about 5.2 × 10−12 dyne-cm from chl/bp = 0 up to chl/bp = 5, whereupon it increases in parallel with the linear DNA. The effect of chloroquine on the secondary structure, torsion constant, and torsional dynamics evidently differs substantially between linear and supercoiled DNAs, even under conditions where the supercoiled DNA is completely relaxed and both DNAs bind the same amount of dye. This strongly contradicts any notion that the local structures of linear and relaxed supercoiled DNA/dye complexes with the same binding ratio are identical. The increase in apparent α at chl/bp = 5 for both DNAs may be due to stacking of the chloroquine in the major groove and consequent stiffening of the filament.