(A) Bacterial two-hybrid between McdA and McdB tagged at their N-termini (X-McdA, X-McdB) or C-termini (McdA-X, McdB-X) on X-gal supplemented plate. (B) Comparison of the ATPase activities of His-MBP-McdA, F SopA-His, and P1 ParA in the presence (solid line, filled symbols) and absence (dashed line, open symbols) of nsDNA. (C) Coelution of ATPase activity with His-MBP-McdA concentration. Purified His-MBP-McdA was further purified over a Superdex 200 column. Protein concentration (bar) and ATPase activity (line) were measured for each fraction indicated. Inset shows an SDS-PAGE gel of the purified fractions. (D) Comparison of ATPase-specific activity of P1 ParA and F SopA-His in the presence (solid line) and absence (dashed line) of nsDNA. (E) Concentration dependency on the specific activity of His-MBP-McdA in the presence (solid line) and absence (dashed line) of nsDNA. (F) ATPase activity assays of McdA-GFP-His with/without DNA and/or McdB-His. (G) Comparison of the ATPase activity of His-MBP-McdA in the presence of increasing McdB concentration with (solid line) or without (dashed line) nsDNA present. Error bars represent SD from at least three independent experiments.