SCF/{beta}-TrCP promotes glycogen synthase kinase 3-dependent degradation of the Nrf2 transcription factor in a Keap1-independent manner

TY - JOUR

T1 - SCF/{beta}-TrCP promotes glycogen synthase kinase 3-dependent degradation of the Nrf2 transcription factor in a Keap1-independent manner

A1 - Rada,Patricia

A1 - Rojo,Ana I.

A1 - Chowdhry,Sudhir

A1 - McMahon,Michael

A1 - Hayes,John D.

A1 - Cuadrado,Antonio

AU - Rada,Patricia

AU - Rojo,Ana I.

AU - Chowdhry,Sudhir

AU - McMahon,Michael

AU - Hayes,John D.

AU - Cuadrado,Antonio

PY - 2011/3

Y1 - 2011/3

N2 - 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/ß-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(?ETGE) 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(?ETGE 6S/6A) protein, lacking these Ser residues, exhibited a longer half-life than Nrf2(?ETGE). Moreover, Nrf2(?ETGE 6S/6A) was insensitive to ß-TrCP regulation and exhibited lower levels of ubiquitination than Nrf2(?ETGE). GSK-3ß enhanced ubiquitination of Nrf2(?ETGE) but not that of Nrf2(?ETGE 6S/6A). The Nrf2(?ETGE) protein but not Nrf2(?ETGE 6S/6A) coimmunoprecipitated with ß-TrCP, and this association was enhanced by GSK-3ß. Our results show for the first time that Nrf2 is targeted by GSK-3 for SCF/ß-TrCP-dependent degradation. We propose a "dual degradation" model to describe the regulation of Nrf2 under different pathophysiological conditions.

AB - 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/ß-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(?ETGE) 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(?ETGE 6S/6A) protein, lacking these Ser residues, exhibited a longer half-life than Nrf2(?ETGE). Moreover, Nrf2(?ETGE 6S/6A) was insensitive to ß-TrCP regulation and exhibited lower levels of ubiquitination than Nrf2(?ETGE). GSK-3ß enhanced ubiquitination of Nrf2(?ETGE) but not that of Nrf2(?ETGE 6S/6A). The Nrf2(?ETGE) protein but not Nrf2(?ETGE 6S/6A) coimmunoprecipitated with ß-TrCP, and this association was enhanced by GSK-3ß. Our results show for the first time that Nrf2 is targeted by GSK-3 for SCF/ß-TrCP-dependent degradation. We propose a "dual degradation" model to describe the regulation of Nrf2 under different pathophysiological conditions.

U2 - 10.1128/MCB.01204-10

DO - 10.1128/MCB.01204-10

M1 - Article

JO - Molecular and Cellular Biology

JF - Molecular and Cellular Biology

SN - 0270-7306

IS - 6

VL - 31

SP - 1121

EP - 1133

ER -