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Sumoylation and desumoylation assays for a chromatin-remodelling complex in vivo and in vitro

External protocol Created on 03 May 2014

Authors

Jung Hwa Kim, Keun Il Kim, and Sung Hee Baek

Summary

Small ubiquitin-like modifier (SUMO) is a small molecule, but has a variety of regulatory functions in cells [1-3]. SUMO modification is involved in transcriptional regulation, subcellular localization, and protein-protein interactions. SUMO conjugation requires sequential E1-dependent activation, E2-dependent conjugation, and E3-dependent ligation steps. Similar to protein phosphorylation, ubiquitination, acetylation, and methylation, SUMO conjugation and deconjugation are a dynamic processes. SUMOylating enzymes add SUMO to protein substrates, whereas deSUMOylating enzymes remove SUMO from SUMO-conjugated proteins. Many transcription factors and coregulators are SUMOylated and SUMOylation of these factors are mainly involved in transcriptiona repression mechanism [4, 5]. SUMOylation and deSUMOylation assays are available to investigate the dynamics of SUMOylation/deSUMOylation process, and provide exciting ways to study transcriptional regulatory mechanisms. This protocol details both in vivo and in vitro SUMOylation/deSUMOylationg assays that provide combined molecular approaches to study cellular functions of chromatin remodelling complexes.

Further details

The protocol was published on Protocol Exchange in 2006. To see the entire protocol, click on the source link.

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