Figure 5
From: Sumo-dependent substrate targeting of the SUMO protease Ulp1
Distinct and separate features of Ulp1 are required for interaction with SUMO. Two-hybrid analysis of various Ulp1(3) truncation mutants (C580S: catalytically inactive; D451N: deleted salt bridge with small ubiquitin-like modifier (SUMO); ts: mutations including S450N in ulp1ts-333) with SUMO/Smt3-BD. The presence of both Smt3 (pOBD2/TRP1) and Ulp1 constructs (pOAD/LEU2) was confirmed by growth on growth media lacking tryptophan and leucine (-T-L). The interaction between Ulp1 constructs and Smt3 is shown as triplicate patches of cells on media lacking adenine (-A). See Figure 3A for a graphic representation of individual constructs.