Plasmid DNA cleavage by topoisomerase IIα
and IIβ in
the presence of quinolones. A: Plasmid DNA was incubated in the absence
(−T) or presence (+T) of topoisomerase IIα (Top2A, top
gel) or β (Top2B, bottom gel). Linearized plasmid (Lin) is shown
in the first lane. Reactions were incubated with 30 or 300 μM
etoposide (Etop) as a positive control. Reactions were performed with
3, 30, or 300 μM of nalidixic acid (NA), moxifloxacin (Moxi),
gemifloxacin (Gemi), ciprofloxacin (Cipro), or levofloxacin (Levo).
Positions of SSB, DSB, and supercoiled (SC) DNA are denoted at left.
Representative gels are shown, and experiments were performed at least
three times. B: quantification of double-strand DNA cleavage relative
to cleavage in the absence of quinolones is shown for both topoisomerase
II isoforms. As a control, etoposide at 30 μM is shown compared
with 300 μM of the quinolones. Dotted line represents the level
of cleavage in the presence of dimethyl sulfoxide (DMSO). Error bars
represent the standard deviation of three or more independent experiments.