Figure 7.

Structurally diverse STIs are able to trap SC and H-SC formed with N155H and Q148H IN. N155H or Q148H IN (30 nM) were pre-assembled with 5′-³²P end-labeled 1.6 kb U5 DNA substrate (0.5 nM) at 14°C for 15 min. Inhibitor and a supercoiled DNA (1.5 nM) were added and the integration reaction was allowed for 3 h at 37°C. Reactions were stopped with 25 mM EDTA and aliquots were subjected to native agarose gel electrophoresis. Panel A and B shows the products obtained with N155H and Q148H IN, respectively. In both panels, lane 1, marked M contains ³²P- end-labeled molecular weight markers. Lane 2, marked C, is control reaction without IN. Lane 3 and 4, marked 0, are with IN but without inhibitor for production of STC. Lanes 5 to 10 contain increasing concentrations of MK-2048, lanes 11–16 contain RAL and lanes 17–22 contain EVG. Trapped SC and H-SC are marked on right. A decrease in formation of STC with increasing concentrations of inhibitor is evident.