Fig. 8
DNA cleavage by Vsr endonucleases in the simultaneous presence of MutLNgo and MutS proteins. The activity of (a) the V.NgoAXIII or (b) V.NgoAXIV endonucleases. The graphs show the data obtained and fitted to a first-order rate equation using Origin 8.5 software. Below the graphs are examples of the electrophoresis profiles of the reaction products resolved in a 10% polyacrylamide gel after digestion of substrate DNA by V.NgoAXIII or V.NgoAXIV: (black triangles) DNA cleavage by gonococcal Vsr endonuclease in the absence of MutLNgo, MutS proteins and 1 mM ATP; (black circles) DNA cleavage by gonococcal Vsr endonuclease in the presence of MutLNgo, MutS proteins, and 1 mM ATP, molar ratio Vsr:MutL:MutS 1:1:1; (black squares) DNA cleavage by gonococcal Vsr endonuclease in the presence of MutLNgo, MutS proteins, and 1 mM ATP, molar ratio Vsr:MutL:MutS 1:2:2. M - GeneRuler 50 bp DNA Ladder (Thermo Scientific). The presented results were obtained using M.NgoA302P-sub (GTCGGT/ACCGGC) for V.NgoAXIII and M.NgoA1175P-sub (CTGG/CCGG) substrate DNA for V.NgoAXIV. Arrows indicate substrate and reaction products obtained after DNA cleavage by a given Vsr endonuclease. P1 and P2 reaction products; S – substrate DNA. The experiments were performed in triplicate, and representative images are shown