Skip to main content
. 2010 Oct 6;285(50):38861–38868. doi: 10.1074/jbc.M110.165191

FIGURE 3.

FIGURE 3.

Dna2 requires a free 5′-end for helicase activity. Helicase activity was assayed as described under “Experimental Procedures.” Reactions containing 5 fmol of either (A) unblocked 53-nt flap substrate (U4:T3:D6); (B) streptavidin blocked 53-nt flap substrate; (C) forked substrate (T3:D6); (D) 5′ overhang substrate (T1:U1) were incubated with either Dna2E675A or Pif1 helicase for 15 min at 37 °C. Lanes 1, 7, 13, 19 are the substrate-alone controls, and lanes 6, 12, 18, 24 are the labeled primer alone controls. Dna2E675A was titrated into the reactions at a concentration of 100 fmol (lanes 2, 8, 14, 20), 250 fmol (lanes 3, 9, 15, 21), and 500 fmol (lanes 4, 10, 16, 22). Lanes 5, 11, 17, 23 contained 500 fmol of Pif 1. The asterisk on the substrates depicted above the figures, denote the position of the radiolabel. Positions of the substrate and helicase products are indicated in the figure.