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SCF/β-TrCP Promotes Glycogen Synthase Kinase 3-Dependent Degradation of the Nrf2 Transcription Factor in a Keap1-Independent Manner
Open accessAbstract
Regulation of transcription factor Nrf2 (NF-E2-related factor 2) involves redox-sensitive proteasomal degradation via the E3 ubiquitin ligase Keap1/Cul3. However, Nrf2 is controlled by other mechanisms that have not yet been elucidated. We now show that glycogen synthase kinase 3 (GSK-3) phosphorylates a group of Ser residues in the Neh6 domain of mouse Nrf2 that overlap with an SCF/β-TrCP destruction motif (DSGIS, residues 334 to 338) and promotes its degradation in a Keap1-independent manner. Nrf2 was stabilized by GSK-3 inhibitors in Keap1-null mouse embryo fibroblasts. Similarly, an Nrf2(ΔETGE) mutant, which cannot be degraded via Keap1, accumulated when GSK-3 activity was blocked. Phosphorylation of a Ser cluster in the Neh6 domain of Nrf2 stimulated its degradation because a mutant Nrf2(ΔETGE 6S/6A) protein, lacking these Ser residues, exhibited a longer half-life than Nrf2(ΔETGE). Moreover, Nrf2(ΔETGE 6S/6A) was insensitive to β-TrCP regulation and exhibited lower levels of ubiquitination than Nrf2(ΔETGE). GSK-3β enhanced ubiquitination of Nrf2(ΔETGE) but not that of Nrf2(ΔETGE 6S/6A). The Nrf2(ΔETGE) protein but not Nrf2(ΔETGE 6S/6A) coimmunoprecipitated with β-TrCP, and this association was enhanced by GSK-3β. Our results show for the first time that Nrf2 is targeted by GSK-3 for SCF/β-TrCP-dependent degradation. We propose a "dual degradation" model to describe the regulation of Nrf2 under different pathophysiological conditions.
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“The Neh6 domain is a redox-insensitive degron that provides Keap1-independent negative Nrf2 regulation. Similar to Neh2, Neh6 contains two highly conserved 334DSGIS338 and 373DSAPGS378 motifs to which the β-transducin repeat-containing protein (βTrCP) binds, and within the DSGIS motif, a phosphorylation site for glycogen synthase kinase-3 (GSK3) that enhances βTrCP activity upon GSK3-mediated ph”
From Nrf2 and Oxidative Stress: A General Overview of Mechanisms and Implications in Human Disease (2022) · cites this paper as [62]
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“These residues are located in the Neh6 domain of NRF2 [27,28].”
BackgroundRojo, A. I., Medina‐Campos, O. N., Rada, P., Zúñiga-Toalá, A., López-Gazcón, A., Espada, S., Pedraza‐Chaverrí, J., & Cuadrado, A. (2011) · Free Radical Biology and Medicine · doi:10.1016/j.freeradbiomed.2011.11.003 · Cited by 198
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Metadata
- Authors
- Rada, P., Rojo, A. I., Chowdhry, S., McMahon, M., Hayes, J. D., & Cuadrado, A.
- Affiliations
- Biomedical Research Networking Center on Neurodegenerative Diseases (ES)
- University of Dundee (GB)
- Corresponding author
- Cuadrado, Antonio
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 2011-03-01
- Volume
- 31
- Issue
- 6
- Pages
- 1121-1133
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC3067901
- OpenAlex
- W2118394016
- Cited by
- 759
- References
- 55
- Citations by year
- 2026: 40 · 2025: 36 · 2024: 53 · 2023: 76 · 2022: 60 · 2021: 71 · 2020: 80 · 2019: 50 · 2018: 47 · 2017: 59 · 2016: 49 · 2015: 51 · 2014: 27 · 2013: 27 · 2012: 21
- FWCI
- 11.31
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Molecular and Cellular Biology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Discovery Research Portal (University of Dundee) (publishedVersion)
- DIGITAL.CSIC (Spanish National Research Council (CSIC)) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics, phytochemicals, and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Ubiquitin and proteasome pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Keywords
- Nrf2, glycogen synthase kinase-3, β-TrCP, protein degradation, serine phosphorylation, ubiquitination, transcription factor regulation, redox signaling
- MeSH terms
- Glycogen Synthase Kinase 3 beta, Kelch-Like ECH-Associated Protein 1, Amino Acid Sequence, Animals, Cell Line, Cytoskeletal Proteins, Humans, Molecular Sequence Data, Mutation, Phosphorylation, Serine, Protein Structure, Tertiary, Glycogen Synthase Kinase 3, Ubiquitin-Protein Ligases, beta-Transducin Repeat-Containing Proteins, Adaptor Proteins, Signal Transducing, NF-E2-Related Factor 2, Mice, Ubiquitination
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