Figure 1 - full size

Figure 1. Figure 1. Genetic analysis of the interface between Ubc9 and RanGAP1. (a) Stereo view of the Ubc9 active site in complex with RanGAP1–SUMO^14 (PDB entry 1Z5S) in ribbon and solid-bond representation, depicting Ubc9 residues selected for genetic analysis. Residues are labeled and hydrogen-bonding interactions indicated by dashed lines. Yellow, SUMO-1; pink, RanGAP1; blue, Ubc9. Images generated with PyMOL^44. (b) Serial dilutions of S. cerevisiae ubc9 cultures bearing wild-type (WT) UBC9 or indicated ubc9 alleles containing single or double point mutations were spotted on YPAD agar and tested for growth at 23 °C (top), 30 °C (middle) and 37 °C (bottom) (Note: D127S allele was expressed under yeast endogenous promoter; see text). (c) Western blotting analysis for Smt3 conjugates (top) or Ubc9 (bottom) in strains containing indicated ubc9 alleles.