Fig. 2: ssDNA efficiently activates the protease activity of PprI.
From: The Deinococcus protease PprI senses DNA damage by directly interacting with single-stranded DNA
a Activation assays showing the ssDNA-enhanced PprI cleavage. DG-DdrO (8 μM) was incubated with DG-PprI (0.1 μM) in the presence of 2 mM MnCl2 in the absence or presence of 35nt ssDNA (0.1 μM) at 37 °C for 30 minutes. b Quantification of cleavage product of (a). Data are mean ± SD from 3 independent experiments, compared with student unpaired Student’s t test (two-sided). c Metal ion preference of DG-PprI cleavage. DG-PprI (0.1 μM) was incubated with DG-DdrO (8 μM) and ssDNA (0.1 μM) in the presence of 2 mM EDTA or 2 mM divalent metal ions (MnCl2, MgCl2, CaCl2, or ZnCl2, respectively) at 37 °C for 30 minutes. d ssDNA activation assays containing various lengths (6-10nt) and concentrations (0.01 or 0.1 μM) of ssDNA using the same reaction conditions as in panel a. e Quantifications of cleavage product by various lengths (5, 10, 20, 30, 40nt) and concentrations of ssDNA using the same reaction conditions as in panel a. Data represent the means of the three replicates, and the bars represent their standard deviations. Source data are provided as a Source Data file.
