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Cleavage–religation equilibrium. Equal concentrations of hTop1p, htop1Thr729Al, htop1Thr729Pro, htop1Thr729Lys or htop1Thr729Glu were incubated with a single 32P-end-labeled DNA fragment in absence or presence of CPT as described in Materials and methods section. After 30 min the reactions were stopped with 0.5% SDS and the cleavage products resolved by urea/PAGE. C indicates no enzyme control. The arrow indicates the position of a high affinity cleavage site.
