Fig. 4 | Nature Communications

Fig. 4

From: RuvC uses dynamic probing of the Holliday junction to achieve sequence specificity and efficient resolution

Fig. 4

Role of Arg76-mediated base pair disruption and base flipping in Tt-RuvC activity. a Schemes of the HJ substrates that were used in measurements of the fluorescence of 2-aminopurine HJ upon binding to Tt-RuvC. The red X indicates 2-aminopurine. b Change in fluorescence after mixing the 2-aminopurine substrates with different Tt-RuvC variants. The results are expressed as the percent change in DNA fluorescence upon the addition of protein. Data from three independent experiments were averaged and plotted for each value. The error bars represent the standard deviation. Dot plots are showing individual data points. c Scheme of HJs that were used to measure the activity of Tt-RuvC (wild-type and R76A) on substrates with abasic sites. The red Y indicates an abasic nucleotide. d, e Resolving activity of the Tt-RuvC variants [wild-type (d) and R76A (e)] that acted on the control and abasic site substrates. Data from four independent experiments were averaged and plotted for each timepoint. Error bars represent the standard deviation

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