The Wnt Pool of Glycogen Synthase Kinase 3β Is Critical for Trophic-Deprivation-Induced Neuronal Death

ABSTRACT Glycogen synthase kinase 3 (GSK-3) is implicated in neuronal death through a causal role, and precise mechanisms have not been unambiguously defined. We show that short hairpin RNA (shRNA) knockdown of GSK-3β, but not GSK-3α, protects cerebellar granule neurons from trophic-deprivation-induced death. Using compartment-targeted inhibitors of the Wnt-regulated GSK-3 pool, NLS-FRAT1, NES-FRAT1, and axin-GSK-3-interacting domain (axin-GID), we locate proapoptotic GSK-3 action to the cytosol and regulation of Bim protein turnover despite constitutive cycling of GSK-3 between the cytosol and nucleus, revealed by leptomycin B. We examine the importance of Ser21/9 (GSK-3α/β) phosphorylation on proapoptotic GSK-3 function. Neurons isolated from GSK-3α/βS21A/S9A knock-in mice survive normally and are fully sensitive to trophic-deprivation-induced death. Nonetheless, inhibition of GSK-3 catalytic activity with lithium or SB216763 protects GSK-3α/βS21A/S9A neurons from death. This indicates that dephosphorylation of GSK-3β/Ser9 and GSK-3α/Ser21 is insufficient for GSK-3 proapoptotic function and that another level of regulation is required. Gel filtration reveals a stress-induced loss of neuronal GSK-3β from a high-molecular-mass complex with a concomitant decrease in axin-bound GSK-3β. These data imply that Wnt-regulated GSK-3β plays a nonredundant role in trophic-deprivation-induced death of neurons.

[1]  D. Mayer,et al.  Glycogen synthase kinase-3 protects estrogen receptor alpha from proteasomal degradation and is required for full transcriptional activity of the receptor. , 2007, Molecular endocrinology.

[2]  D. Chuang,et al.  Differential Roles of Glycogen Synthase Kinase-3 Isoforms in the Regulation of Transcriptional Activation* , 2006, Journal of Biological Chemistry.

[3]  J. Lucas,et al.  Full Reversal of Alzheimer's Disease-Like Phenotype in a Mouse Model with Conditional Overexpression of Glycogen Synthase Kinase-3 , 2006, The Journal of Neuroscience.

[4]  K. Cadigan,et al.  Wnt signaling: complexity at the surface , 2006, Journal of Cell Science.

[5]  A. Strasser,et al.  Mutually Exclusive Subsets of BH3-Only Proteins Are Activated by the p53 and c-Jun N-Terminal Kinase/c-Jun Signaling Pathways during Cortical Neuron Apoptosis Induced by Arsenite , 2005, Molecular and Cellular Biology.

[6]  J. Filén,et al.  Constitutively Active Cytoplasmic c-Jun N-Terminal Kinase 1 Is a Dominant Regulator of Dendritic Architecture: Role of Microtubule-Associated Protein 2 as an Effector , 2005, The Journal of Neuroscience.

[7]  H. Manji,et al.  Glycogen Synthase Kinase-3: a Putative Molecular Target for Lithium Mimetic Drugs , 2005, Neuropsychopharmacology.

[8]  D. Alessi,et al.  Role that phosphorylation of GSK3 plays in insulin and Wnt signalling defined by knockin analysis , 2005, The EMBO journal.

[9]  Ming-tao Li,et al.  Activity deprivation-dependent induction of the proapoptotic BH3-only protein Bim is independent of JNK/c-Jun activation during apoptosis in cerebellar granule neurons , 2005, Neuroscience Letters.

[10]  P. Coffer,et al.  Cytokine mediated suppression of TF‐1 apoptosis requires PI3K activation and inhibition of Bim expression , 2005, FEBS letters.

[11]  Esther B. E. Becker,et al.  Characterization of the c-Jun N-Terminal Kinase-BimEL Signaling Pathway in Neuronal Apoptosis , 2004, The Journal of Neuroscience.

[12]  P. Greengard,et al.  Pharmacological inhibitors of glycogen synthase kinase 3. , 2004, Trends in pharmacological sciences.

[13]  Jiahuai Han,et al.  Distinct Requirements for p38α and c-Jun N-terminal Kinase Stress-activated Protein Kinases in Different Forms of Apoptotic Neuronal Death* , 2004, Journal of Biological Chemistry.

[14]  J. Woodgett,et al.  Glycogen Synthase Kinase-3β Haploinsufficiency Mimics the Behavioral and Molecular Effects of Lithium , 2004, The Journal of Neuroscience.

[15]  E. Aronica,et al.  Induction of Dickkopf-1, a Negative Modulator of the Wnt Pathway, Is Associated with Neuronal Degeneration in Alzheimer's Brain , 2004, The Journal of Neuroscience.

[16]  R. Jope,et al.  Glycogen synthase kinase-3&bgr; is highly activated in nuclei and mitochondria , 2003, Neuroreport.

[17]  J. Woodgett,et al.  Physiological roles of glycogen synthase kinase-3: potential as a therapeutic target for diabetes and other disorders. , 2003, Current drug targets. Immune, endocrine and metabolic disorders.

[18]  T. Herdegen,et al.  Lithium Blocks the c-Jun Stress Response and Protects Neurons via Its Action on Glycogen Synthase Kinase 3 , 2003, Molecular and Cellular Biology.

[19]  Eugene M. Johnson,et al.  JNK-Mediated BIM Phosphorylation Potentiates BAX-Dependent Apoptosis , 2003, Neuron.

[20]  Christina A. Wilson,et al.  GSK-3α regulates production of Alzheimer's disease amyloid-β peptides , 2003, Nature.

[21]  D. Chuang,et al.  Postinsult treatment with lithium reduces brain damage and facilitates neurological recovery in a rat ischemia/reperfusion model , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Minoru Yoshida,et al.  Nucleo-cytoplasmic transport of proteins as a target for therapeutic drugs. , 2003, Current medicinal chemistry.

[23]  B. Doble,et al.  GSK-3: tricks of the trade for a multi-tasking kinase , 2003, Journal of Cell Science.

[24]  S. DeRuiter,et al.  Simultaneous inhibition of GSK3α and GSK3β using hairpin siRNA expression vectors , 2003 .

[25]  T. Dale,et al.  The Regulation of Glycogen Synthase Kinase-3 Nuclear Export by Frat/GBP* , 2002, The Journal of Biological Chemistry.

[26]  T. Herdegen,et al.  c-Jun N-Terminal Protein Kinase (JNK) 2/3 Is Specifically Activated by Stress, Mediating c-Jun Activation, in the Presence of Constitutive JNK1 Activity in Cerebellar Neurons , 2002, The Journal of Neuroscience.

[27]  R. Jope,et al.  The multifaceted roles of glycogen synthase kinase 3β in cellular signaling , 2001, Progress in Neurobiology.

[28]  R. Jope,et al.  Proapoptotic Stimuli Induce Nuclear Accumulation of Glycogen Synthase Kinase-3β* , 2001, The Journal of Biological Chemistry.

[29]  P. Cohen,et al.  The renaissance of GSK3 , 2001, Nature Reviews Molecular Cell Biology.

[30]  J. Woodgett,et al.  Glycogen synthase kinase-3: properties, functions, and regulation. , 2001, Chemical reviews.

[31]  Laurence H. Pearl,et al.  Crystal Structure of Glycogen Synthase Kinase 3β Structural Basis for Phosphate-Primed Substrate Specificity and Autoinhibition , 2001, Cell.

[32]  P. Cohen,et al.  A common phosphate binding site explains the unique substrate specificity of GSK3 and its inactivation by phosphorylation. , 2001, Molecular Cell.

[33]  A. Reith,et al.  Selective small‐molecule inhibitors of glycogen synthase kinase‐3 activity protect primary neurones from death , 2001, Journal of neurochemistry.

[34]  O. Bernard,et al.  Dominant-Negative c-Jun Promotes Neuronal Survival by Reducing BIM Expression and Inhibiting Mitochondrial Cytochrome c Release , 2001, Neuron.

[35]  Ming-tao Li,et al.  Cyclic AMP Promotes Neuronal Survival by Phosphorylation of Glycogen Synthase Kinase 3β , 2000, Molecular and Cellular Biology.

[36]  F. McCormick,et al.  Differential Regulation of Glycogen Synthase Kinase 3β by Insulin and Wnt Signaling* , 2000, The Journal of Biological Chemistry.

[37]  M. Dickens,et al.  Dual Roles for c-Jun N-Terminal Kinase in Developmental and Stress Responses in Cerebellar Granule Neurons , 2000, The Journal of Neuroscience.

[38]  J W Yates,et al.  Selective small molecule inhibitors of glycogen synthase kinase-3 modulate glycogen metabolism and gene transcription. , 2000, Chemistry & biology.

[39]  Eugene M. Johnson,et al.  Phosphatidylinositol 3-Kinase Is Required for the Trophic, But Not the Survival-Promoting, Actions of NGF on Sympathetic Neurons , 2000, The Journal of Neuroscience.

[40]  J. Woodgett,et al.  Requirement for glycogen synthase kinase-3β in cell survival and NF-κB activation , 2000, Nature.

[41]  P. Cohen,et al.  A GSK3‐binding peptide from FRAT1 selectively inhibits the GSK3‐catalysed phosphorylation of Axin and β‐catenin , 1999, FEBS letters.

[42]  M. Courtney,et al.  The mechanism of Ara‐C‐induced apoptosis of differentiating cerebellar granule neurons , 1999, The European journal of neuroscience.

[43]  M. Mercken,et al.  Presenilin 1 associates with glycogen synthase kinase-3beta and its substrate tau. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M. Yanagida,et al.  Leptomycin B inhibition of signal-mediated nuclear export by direct binding to CRM1. , 1998, Experimental cell research.

[45]  L. Rubin,et al.  Phosphorylation of c-Jun Is Necessary for Apoptosis Induced by Survival Signal Withdrawal in Cerebellar Granule Neurons , 1998, The Journal of Neuroscience.

[46]  J. Woodgett,et al.  Molecular cloning and expression of glycogen synthase kinase‐3/factor A. , 1990, The EMBO journal.

[47]  M. Cambray-Deakin,et al.  The cellular neurobiology of neuronal development: The cerebellar granule cell , 1988, Brain Research Reviews.

[48]  T. Laessig,et al.  Glycogen synthase kinase-3beta phosphorylates Bax and promotes its mitochondrial localization during neuronal apoptosis. , 2004, Journal of Neuroscience.

[49]  Christina A. Wilson,et al.  GSK-3alpha regulates production of Alzheimer's disease amyloid-beta peptides. , 2003, Nature.

[50]  S. DeRuiter,et al.  Simultaneous inhibition of GSK3alpha and GSK3beta using hairpin siRNA expression vectors. , 2003, Molecular therapy : the journal of the American Society of Gene Therapy.

[51]  J. Woodgett,et al.  Requirement for glycogen synthase kinase-3beta in cell survival and NF-kappaB activation. , 2000, Nature.

[52]  M. Hetman,et al.  Role of glycogen synthase kinase-3beta in neuronal apoptosis induced by trophic withdrawal. , 2000, The Journal of neuroscience : the official journal of the Society for Neuroscience.