Protocol to modulate SUMOylation of a specific protein in budding yeast using chemical genetic approaches
Year: 2025
Type: article
Abstract: 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 • Complete and user-friendly protocol for altering SUMOylation of a specific protein • Steps for transforming yeast cells with plasmids expressing SUMO pathway enzymes • Procedures for the expression of SUMO enzymes from plasmids and SUMOylation analysis Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.
Source: STAR Protocols
Authors: Emily Gutierrez-Morton, Yanchang Wang
Institution: Florida State University
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
Awards: G344921309, G669161380
OpenAlex ID: W4408164676