Figure 5.

DNA nicking by the pCU1 TraI relaxase in the presence of metals. To determine the DNA nicking activity of the pCU1 TraI relaxase (WT_299) after treatment with EDTA and in the presence of the indicated concentration of metal, 5 μM enzyme was incubated with 1 μM substrate (35/7oriT-hairpin) for 1 h at 37°C, and products were then separated on denaturing polyacrylamide gels. Nicking and religation activity is represented as percent product generated. Each experiment was performed in triplicate; the error is the standard error of these replicates. Additional details can be found in the Materials and Methods section and the Supplementary Data.