Structural and Functional Characterization of Nrf2 Degradation by the Glycogen Synthase Kinase 3/β-TrCP Axis
| Autor: | Patricia Rada, Ana I. Rojo, Nathalie Evrard-Todeschi, Nadia G. Innamorato, Axelle Cotte, Tomasz Jaworski, Julio C. Tobón-Velasco, Herman Devijver, María Flor García-Mayoral, Fred Van Leuven, John D. Hayes, Gildas Bertho, Antonio Cuadrado |
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| Přispěvatelé: | UAM. Departamento de Bioquímica |
| Rok vydání: | 2012 |
| Předmět: |
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Models Molecular Cell signaling NF-E2-Related Factor 2 Medicina Beta-Transducin Repeat-Containing Proteins Molecular Sequence Data environment and public health transcription factor Nrf2 Mice Glycogen Synthase Kinase 3 GSK-3 Serine Animals Humans Amino Acid Sequence Phosphorylation Molecular Biology Peptide sequence Transcription factor Cells Cultured Mice Knockout Glycogen Synthase Kinase 3 beta biology Articles Cell Biology respiratory system beta-Transducin Repeat-Containing Proteins Protein Structure Tertiary Ubiquitin ligase Mice Inbred C57BL HEK293 Cells Biochemistry Mutagenesis Mutagenesis Site-Directed biology.protein Signal transduction |
| Zdroj: | Biblos-e Archivo. Repositorio Institucional de la UAM instname |
| ISSN: | 1098-5549 2010-1872 |
| Popis: | The transcription factor NF-E2-related factor 2 (Nrf2) is a master regulator of a genetic program, termed the phase 2 response, that controls redox homeostasis and participates in multiple aspects of physiology and pathology. Nrf2 protein stability is regulated by two E3 ubiquitin ligase adaptors, Keap1 and β-TrCP, the latter of which was only recently reported. Here, two-dimensional (2D) gel electrophoresis and site-directed mutagenesis allowed us to identify two serines of Nrf2 that are phosphorylated by glycogen synthase kinase 3β (GSK-3β) in the sequence DSGISL. Nuclear magnetic resonance studies defined key residues of this phosphosequence involved in docking to the WD40 propeller of β-TrCP, through electrostatic and hydrophobic interactions. We also identified three arginine residues of β-TrCP that participate in Nrf2 docking. Intraperitoneal injection of the GSK-3 inhibitor SB216763 led to increased Nrf2 and heme oxygenase-1 levels in liver and hippocampus. Moreover, mice with hippocampal absence of GSK-3βexhibited increased levels of Nrf2 and phase 2 gene products, reduced glutathione, and decreased levels of carbonylated proteins and malondialdehyde. This study establishes the structural parameters of the interaction of Nrf2 with the GSK-3/β-TrCP axis and its functional relevance in the regulation of Nrf2 by the signaling pathways that impinge on GSK-3 This work was supported by MICINN grant SAF2010-18722 from the Spanish Ministry of Science and Innovation. Patricia Rada is contracted under the Formación de Profesorado Universitario program of the Spanish Ministry of Science and Innovation. We acknowledge funding and infrastructural support from EEC 7th Framework Program, KU Leuven Research Fund, and KU Leuven Research & Development |
| Databáze: | OpenAIRE |
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