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. 2019 Oct 29;8:e46096. doi: 10.7554/eLife.46096

Figure 4. Single turnover DNA unwinding.

(A) DNA unwinding of equal arm (in blue boxes -■-, 5’-Cy3 labelled at the duplex), 3’-long arm (in green lower left triangle -◣-, 5’-Cy3 labelled at the duplex), 5’-long arm (in red lower right triangle -SsoMCM WT. Experiments were simultaneously initiated with ATP and an unlabelled ssDNA trap oligo identical to the labelled strand as described in the Materials and methods. (B) DNA unwinding of equal arm fork DNA substrate (5’-Cy3 labelled at duplex) with SsoMCM WT (in closed boxes -■-) and –WH (in open boxes -□-). There is an increase in total DNA unwound with –WH compared to WT (grey arrow). (C) DNA unwinding of 3’-long arm fork DNA substrate (5’-Cy3 labelled at duplex) with SsoMCM WT (in lower left closed triangle -◣-) and –WH (in lower left open triangle -◺ -). There is a decrease in the total DNA unwound with –WH compared to WT (grey arrow). Error bars represent standard error from 3 to 5 independent experiments and data was fit to Equation 3.

Figure 4.

Figure 4—figure supplement 1. Single turnover DNA Unwinding.

Figure 4—figure supplement 1.

DNA unwinding of 3’-long arm/5’-arm (n), (n = 0, in green open lower left triangle -◣-, n = 8 in green closed lower left triangle -◣-,5’-Cy3 labelled at the duplex), 5’-long arm/3’-arm (n), (n = 0, in red open lower right triangle -◢-, n = 8 in red open lower right triangle-◢-, 3’-Cy5 labelled at the duplex) fork DNA substrates with SsoMCM WT. Experiments were simultaneously initiated with ATP and an unlabelled ssDNA trap oligo identical to the labelled strand as described in the Materials and methods. There is a decrease in total DNA unwound with n = 0 compared to n = 8 in 3’-long arm/5’-arm(n) fork substrate (grey arrow). Error bars represent standard error from 3 to 5 independent experiments and data was fit to Equation 3.