Fig. 3: Characterization of SLFN11 phosphomimetic mutants S219D, T230D, and S753D.

a Positions of phosphorylation sites S219 and T230 (purple) in the nuclease domains of SLFN11^wt bound to tRNA-Leu. The nuclease active sites of both SLFN11^wt protomers are indicated. b Position of phosphorylation site S753 (purple) in the ssDNA-bound helicase domain of SLFN11^wt (PDB: 7ZES). c MST measurements of SLFN11^wt and phosphomimetic mutants (S219D, T230D, S753D) binding to fluorescently labeled type II tRNA-Leu to determine equilibrium dissociation constants (K_d). The indicated SLFN11 concentrations assume a dimeric state. Data are represented as mean values +/− SD from three independent experiments. d Analysis of tRNA-Leu cleavage by SLFN11^wt, SLFN11^S219D, SLFN11^T230D, and SLFN11^S753D. Data are represented as mean values +/− SD from three independent experiments. e NanoDSF measurements of SLFN11^S753D (left) and SLFN11^wt (right) in the presence of ssDNA and dsDNA. Data are represented as mean values +/− SD from three independent experiments. One representative replicate is shown. f ssDNA binding of SLFN11^wt and SLFN11^S753D (left), as well as SLFN11^S219D and SLFN11^T230D (right) monitored by electrophoretic mobility shift assays. The experiment was performed in duplicates. One representative replicate is shown. Source data for c–f are provided as a Source Data file.