Protocol to modulate SUMOylation of a specific protein in budding yeast using chemical genetic approaches

SUMOylation (small ubiquitin-like modifier) is a ubiquitous and highly dynamic posttranslational modification. Here, we present a protocol to alter the local SUMOylation landscape of target proteins in budding yeast Saccharomyces cerevisiae using chemical genetic tools. We describe steps for recruiting SUMO enzymes (Ulp1^PD or Ubc9) to GFP-tagged proteins using GBP (GFP-binding protein)-fusion proteins. We then detail procedures for inducing SUMO conjugation/deconjugation and the subsequent SUMOylation analysis. For complete details on the use and execution of this protocol, please refer to Gutierrez-Morton et al.¹.