Figure 5. Human ENDOV binds branched DNA substrates.
The affinities of E. coli and human endonuclease V for different DNA substrates were tested by electrophoretic mobility shift assay. Substrates used were: (A): Undamaged DNA, (B): Hypoxanthine, (C): 5′-flap, (D): 3′-flap, (E): pseudo-Y, (F): fork, (G): 3-way junction and (H): Holliday junction (asterisk indicates the 32P-labelled strands). 2–8 pmol E. coli endoV and 1.6–6.4 pmol ENDOV enzymes were assayed with 10 fmol substrates as indicated. All experiments were repeated 2 to 3 times and a representative assay is shown. Bound substrates relative to free were quantified and are shown to the right. F = free DNA, B = bound DNA, - = no enzyme added, filled circles = E. coli endoV; open circles = ENDOV.