Back to article: Identification of SUMO conjugation sites in the budding yeast proteome


FIGURE 3: Mutational analysis of SUMO sites identified. (A) Histidine pulldowns from cells carrying TFG1, RAD16 and FOB1 tagged with 3 HA epitopes in strains expressing SMT3 tagged with 6 histidines (6his-SMT3) or wild-type SMT3 (untagged control). Western blot analysis using α-HA antibodies in the pulldown demonstrates the presence of SUMO conjugates for TFG1, RAD16 and FOB1. (B) Histidine pulldowns from cells carrying RPC53 or RPC53 (K38,115,237,322R) tagged with 3 HA epitopes. The analysis demonstrates that lysines 38, 115, 237 and 322 (identified in our screen) are sites for the conjugation of SMT3 in RPC53. (C) Histidine pulldowns from cells carrying NTG1 or NTG1 (K38, 396R) tagged with 3HA epitopes. The analysis demonstrates that lysines 38 and 396 (identified in our screen) are sites for the conjugation of SMT3 in NTG1.