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In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
Coordinated sumoylation and ubiquitination modulate EGF induced EGR1 expression and stability
Arcangela Gabriella Manente1, Giulia Pinton, Daniela Tavian
1Dipartimento di Scienze Chimiche, Alimentari, Farmaceutiche e Farmacologiche, University of Piemonte Orientale "A. Avogadro", Novara, Italy.
Plos One
|October 15, 2011
Summary
Sumoylation of early growth response-1 (EGR1) protein influences its stability. EGF stimulation enhances EGR1 ubiquitination and degradation through SUMO-1 modification, impacting gene transcription.
Area of Science:
- Molecular Biology
- Cell Signaling
- Transcription Factors
Background:
- Early growth response-1 (EGR1) is a transcription factor induced by epidermal growth factor (EGF).
- EGR1 expression is regulated by signaling pathways like Erk1/2 and PI3K/Akt/Forkhead.
- Post-translational modifications, including sumoylation, affect EGR1 activity and stability.
Purpose of the Study:
- To investigate the role of sumoylation in EGF-induced Egr1 expression and stability.
- To elucidate the impact of SUMO-1 modification on EGR1 turnover.
Main Methods:
- Modulation of sumoylation proteins in ECV304 cells via transient transfection.
- Evaluation of Egr1 expression (mRNA and protein) following EGF treatment.
- Analysis of protein ubiquitination and degradation pathways.
Main Results:
- EGF treatment induced transient Egr1 expression, with a fraction of endogenous EGR1 being sumoylated.
- Overexpression of SUMO-1/Ubc9 stabilized EGF-induced ERK1/2 phosphorylation and increased Egr1 transcription.
- Conversely, SUMO-1/Ubc9 overexpression reduced EGR1 protein levels, suggesting enhanced degradation.
Conclusions:
- SUMO-1 modification enhances EGR1 ubiquitination upon EGF stimulation.
- This process is involved in modulating EGR1 stability and turnover via proteasomal degradation.
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