Fig. 2: Role of a C-terminal SIM motif (SIM2) in the catalytic activity of Nse2.
From: Structural basis for the E3 ligase activity enhancement of yeast Nse2 by SUMO-interacting motifs
a Multiple alignment of Nse2 sequences from Saccharomyces cerevisiae, S. eubayanus, S. paradoxus, Zygos. parabailli, Schizos. pombe, Mus musculus and Homo sapiens (in red SIM2). b Electron density maps of SIM2 Nse2 and SUMOB. c. Stick representation of the complex between SIM2 and SUMOB. d Structural alignment of Nse2 SIM2 and SIM2 from PIAS1-peptide complex (PDB 6V7P), evidencing similar locations of Ile264 and Val266 in the SUMOB cavity. e–g Multiple-turnover SUMOylation reactions of yeast Nse2/Arm-Smc5 complex (wild type and mutants) using cP53, cNse4 and coil/Smc5 substrates. Enzyme concentration was chosen based on the concentration dependence assay (left). Error bars showing that point mutations or deletion of SIM2 reduce E3 activity (right). Data values represent the mean ±SD, n = 3 technical replicates. Significance was measured by a two-tailed unpaired t-test relative to wild type. All data were analyzed with a 95% confidence interval. *P < 0.05, **P < 0.01, ***P < 0.001. Exact P values from the left to right: (e) 0.027, 0.009, 0.002; (f) 0.0003, 0.0003, 0.0001; (g) 0.021, 0.002, 0.002. Source data are provided as a Source Data file.
