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 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..
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