An Inverse Agonist GSK5182 Increases Protein Stability of the Orphan Nuclear Receptor ERRγ via Inhibition of Ubiquitination
Abstract
:1. Introduction
2. Results
2.1. An Inverse Agonist, GSK5182, Stabilizes the ERRγ Protein
2.2. Different Effect of Ligands on ERRγ Protein
2.3. Protein Stability Due to GSK5182 Requires Y326 of ERRγ
2.4. GSK5182 Stabilizes ERRγ by Extending Its Half-Life
2.5. GSK5182 Prevents ERRγ Ubiquitination by Inhibiting Its Association with the E3 Ligase Parkin
2.6. GSK5182 Promotes Recruitment of the Corepressor SMILE to ERRγ
2.7. The AF-2 Domain of ERRγ Is Crucial for Protein Turnover
3. Discussion
4. Materials and Methods
4.1. Chemicals
4.2. Plasmids, DNA Constructs, and Recombinant Adenoviruses
4.3. Cell Culture, Transient Transfection, and Luciferase Assay
4.4. Animal Experiments
4.5. Chromatin Immunoprecipitation (ChIP) Assay
4.6. Pulse Labeling
4.7. Confocal Microscopy
4.8. Immunoblotting
4.9. Nuclear/Cytosol Fractionation
4.10. Statistical Analyses
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Na, S.-Y.; Kim, K.-S.; Jung, Y.S.; Kim, D.-K.; Kim, J.; Cho, S.J.; Lee, I.-K.; Chung, J.; Kim, J.-S.; Choi, H.-S. An Inverse Agonist GSK5182 Increases Protein Stability of the Orphan Nuclear Receptor ERRγ via Inhibition of Ubiquitination. Int. J. Mol. Sci. 2023, 24, 96. https://doi.org/10.3390/ijms24010096
Na S-Y, Kim K-S, Jung YS, Kim D-K, Kim J, Cho SJ, Lee I-K, Chung J, Kim J-S, Choi H-S. An Inverse Agonist GSK5182 Increases Protein Stability of the Orphan Nuclear Receptor ERRγ via Inhibition of Ubiquitination. International Journal of Molecular Sciences. 2023; 24(1):96. https://doi.org/10.3390/ijms24010096
Chicago/Turabian StyleNa, Soon-Young, Ki-Sun Kim, Yoon Seok Jung, Don-Kyu Kim, Jina Kim, Sung Jin Cho, In-Kyu Lee, Jongkyeong Chung, Jeong-Sun Kim, and Hueng-Sik Choi. 2023. "An Inverse Agonist GSK5182 Increases Protein Stability of the Orphan Nuclear Receptor ERRγ via Inhibition of Ubiquitination" International Journal of Molecular Sciences 24, no. 1: 96. https://doi.org/10.3390/ijms24010096