Structural and functional characterization of Nrf2 degradation by the glycogen synthase kinase 3/β-TrCP Axis

TY - JOUR

T1 - Structural and functional characterization of Nrf2 degradation by the glycogen synthase kinase 3/β-TrCP Axis

A1 - Rada,Patricia

A1 - Rojo,Ana I.

A1 - Evrard-Todeschi,Nathalie

A1 - Innamorato,Nadia G.

A1 - Cotte,Axelle

A1 - Jaworski,Tomasz

A1 - Tobón-Velasco,Julio C.

A1 - Devijver,Herman

A1 - García-Mayoral,María Flor

A1 - Van Leuven,Fred

A1 - Hayes,John D.

A1 - Bertho,Gildas

A1 - Cuadrado,Antonio

AU - Rada,Patricia

AU - Rojo,Ana I.

AU - Evrard-Todeschi,Nathalie

AU - Innamorato,Nadia G.

AU - Cotte,Axelle

AU - Jaworski,Tomasz

AU - Tobón-Velasco,Julio C.

AU - Devijver,Herman

AU - García-Mayoral,María Flor

AU - Van Leuven,Fred

AU - Hayes,John D.

AU - Bertho,Gildas

AU - Cuadrado,Antonio

PY - 2012

Y1 - 2012

N2 - 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 ß-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 3ß (GSK-3ß) in the sequence DSGISL. Nuclear magnetic resonance studies defined key residues of this phosphosequence involved in docking to the WD40 propeller of ß-TrCP, through electrostatic and hydrophobic interactions. We also identified three arginine residues of ß-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-3ß exhibited 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/ß-TrCP axis and its functional relevance in the regulation of Nrf2 by the signaling pathways that impinge on GSK-3.

AB - 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 ß-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 3ß (GSK-3ß) in the sequence DSGISL. Nuclear magnetic resonance studies defined key residues of this phosphosequence involved in docking to the WD40 propeller of ß-TrCP, through electrostatic and hydrophobic interactions. We also identified three arginine residues of ß-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-3ß exhibited 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/ß-TrCP axis and its functional relevance in the regulation of Nrf2 by the signaling pathways that impinge on GSK-3.

U2 - 10.1128/MCB.00180-12

DO - 10.1128/MCB.00180-12

M1 - Article

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

IS - 17

VL - 32

SP - 3486

EP - 3499

ER -