Related Experiment Video
Updated: May 30, 2026
09:45
In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Published on: January 29, 2018
Analysis of androgen receptor SUMOylation
Miia M Rytinki1, Sanna Kaikkonen, Päivi Sutinen
1Institute of Biomedicine, University of Eastern Finland, Kuopio, Finland. miia.rytinki@uef.fi
Methods in Molecular Biology (Clifton, N.J.)
|July 29, 2011
Summary
Androgen receptor (AR) SUMOylation, a post-translational modification, represses its transcriptional activity in prostate cancer. Protocols are detailed for detecting this crucial AR modification in cells.
Area of Science:
- Molecular Biology
- Cell Biology
- Biochemistry
Background:
- Androgen receptor (AR) is a key transcription factor in prostate cancer.
- AR activity is regulated by post-translational modifications (PTMs), including SUMOylation.
- SUMOylation involves the conjugation of small ubiquitin-related modifier (SUMO) proteins.
Purpose of the Study:
- To investigate the SUMOylation of the Androgen Receptor (AR).
- To characterize the functional impact of AR SUMOylation on its transcriptional activity.
- To describe methodologies for analyzing AR SUMOylation in cultured cells.
Main Methods:
- Immunoblotting
- Gel mobility shift assays
- Immunoprecipitation
- Analysis of AR SUMOylation in cultured cells
Main Results:
- AR is modified by SUMO-1 at two conserved lysine residues in its N-terminal domain.
- This SUMOylation is agonist-enhanced and represses AR transcriptional activity.
- The repression is reversible and target gene-selective.
- Cellular SUMOylation levels are typically low, often requiring overexpression for detection.
Conclusions:
- SUMOylation is a significant post-translational modification regulating AR function.
- Specific protocols are established for detecting and analyzing AR SUMOylation.
- These methods are broadly applicable to studying SUMOylation of other nuclear receptors.
More Related Videos
Related Concept Videos
02:04Covalently Linked Protein Regulators
Proteins can undergo many types of post-translational modifications, often in response to changes in their environment. These modifications play an important role in the function and stability of these proteins. Covalently linked molecules include functional groups, such as methyl, acetyl, and phosphate groups, and also small proteins, such as ubiquitin. There are around 200 different types of covalent regulators that have been identified.
These groups modify specific amino acids in a protein.
These groups modify specific amino acids in a protein.
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...