Figure 6. Mdm2 sumoylates p53 in yeast on lysine 386 and sumoylation is required for p53-Mdm2 complexes recruitment to yeast nuclear bodies.

(A) His-tagged proteins were purified from cell extracts and analyzed by immunoblotting using anti-p53 and anti-Smt3 antibodies. (B,C) Sumoylated p53 (asterisks) as detected by immunoblotting on crude yeast lysates. An irrelevant lane was cut off as indicated by a space. PGK immunoblotting served as loading control. (D–G) Maximum projections from fluorescence image stacks of live yeast cells expressing the indicated fusion proteins were obtained with a DeltaVision workstation. Bar, 2 µm.