The PEA-15 Protein Regulates Autophagy via Activation of JNK*

PEA-15/PED (phosphoprotein enriched in astrocytes 15 kDa/phosphoprotein enriched in diabetes) is a death effector domain-containing protein which is known to modulate apoptotic cell death. The mechanism by which PEA-15 inhibits caspase activation and increases ERK (extracellular-regulated kinase) activity is well characterized. Here, we demonstrate that PEA-15 is not only pivotal in the activation of the ERK pathway but also modulates JNK (c-Jun N-terminal kinase) signaling. Upon overexpression of PEA-15 in malignant glioma cells, JNK is potently activated. The PEA-15-induced JNK activation depends on the phosphorylation of PEA-15 at both phosphorylation sites (serine 104 and serine 116). The activation of JNK is substantially inhibited by siRNA-mediated down-regulation of endogenous PEA-15. Moreover, we demonstrate that glioma cells overexpressing PEA-15 show increased signs of autophagy in response to classical autophagic stimuli such as ionizing irradiation, serum deprivation, or rapamycin treatment. In contrast, the non-phosphorylatable mutants of PEA-15 are not capable of promoting autophagy. The inhibition of JNK abrogates the PEA-15-mediated increase in autophagy. In conclusion, our data show that PEA-15 promotes autophagy in glioma cells in a JNK-dependent manner. This might render glioma cells more resistant to adverse stimuli such as starvation or ionizing irradiation.

[1]  T. Mikkelsen,et al.  The induction of autophagy by γ‐radiation contributes to the radioresistance of glioma stem cells , 2009, International journal of cancer.

[2]  A. Thorburn,et al.  Regulation of HMGB1 release by autophagy , 2009, Autophagy.

[3]  M. Gleave,et al.  PTEN Loss Promotes Mitochondrially Dependent Type II Fas-Induced Apoptosis via PEA-15 , 2008, Molecular and Cellular Biology.

[4]  Yan Cheng,et al.  ERK and JNK mediate TNFalpha-induced p53 activation in apoptotic and autophagic L929 cell death. , 2008, Biochemical and biophysical research communications.

[5]  F. Yamasaki,et al.  PEA-15 induces autophagy in human ovarian cancer cells and is associated with prolonged overall survival. , 2008, Cancer research.

[6]  S. Pattingre,et al.  JNK1-mediated phosphorylation of Bcl-2 regulates starvation-induced autophagy. , 2008, Molecular cell.

[7]  P. Codogno,et al.  Role of non-canonical Beclin 1-independent autophagy in cell death induced by resveratrol in human breast cancer cells , 2008, Cell Death and Differentiation.

[8]  M. Hollstein,et al.  The PEA-15/PED protein protects glioblastoma cells from glucose deprivation-induced apoptosis via the ERK/MAP kinase pathway , 2008, Oncogene.

[9]  G. Fuller,et al.  Constitutive activation of c-Jun N-terminal kinase correlates with histologic grade and EGFR expression in diffuse gliomas , 2008, Journal of Neuro-Oncology.

[10]  T. Ikejima,et al.  Oridonin induced autophagy in human cervical carcinoma HeLa cells through Ras, JNK, and P38 regulation. , 2007, Journal of pharmacological sciences.

[11]  S. Krupenko,et al.  Cooperation between JNK1 and JNK2 in activation of p53 apoptotic pathway , 2007, Oncogene.

[12]  Y. Atomi,et al.  Autophagy is activated in colorectal cancer cells and contributes to the tolerance to nutrient deprivation. , 2007, Cancer research.

[13]  B. Levine,et al.  Cell biology: Autophagy and cancer , 2007, Nature.

[14]  Per O. Seglen,et al.  Methods for Monitoring Autophagy from Yeast to Human , 2007, Autophagy.

[15]  Shuang-yin Han,et al.  c-Jun NH2-Terminal Kinase 2α2 Promotes the Tumorigenicity of Human Glioblastoma Cells , 2006 .

[16]  F. Beuvon,et al.  Phosphoprotein enriched in astrocytes-15 kDa expression inhibits astrocyte migration by a protein kinase C delta-dependent mechanism. , 2006, Molecular biology of the cell.

[17]  H. Saya,et al.  Antitumor effect of E1A in ovarian cancer by cytoplasmic sequestration of activated ERK by PEA15 , 2006, Oncogene.

[18]  G. Troncone,et al.  Raised expression of the antiapoptotic protein ped/pea-15 increases susceptibility to chemically induced skin tumor development , 2005, Oncogene.

[19]  Michael D. Schneider,et al.  Bcl-2 Antiapoptotic Proteins Inhibit Beclin 1-Dependent Autophagy , 2005, Cell.

[20]  Eric H. Baehrecke,et al.  Autophagy: dual roles in life and death? , 2005, Nature Reviews Molecular Cell Biology.

[21]  Y. Kondo,et al.  Inhibition of the DNA-dependent protein kinase catalytic subunit radiosensitizes malignant glioma cells by inducing autophagy. , 2005, Cancer research.

[22]  Y. Kondo,et al.  Radiation-induced autophagy is associated with LC3 and its inhibition sensitizes malignant glioma cells. , 2005, International journal of oncology.

[23]  C. Soeller,et al.  C-terminal splicing of NTPDase2 provides distinctive catalytic properties, cellular distribution and enzyme regulation. , 2005, The Biochemical journal.

[24]  Sam-Yong Park,et al.  Structural basis for the selective inhibition of JNK1 by the scaffolding protein JIP1 and SP600125 , 2004, The EMBO journal.

[25]  Daniel J Klionsky,et al.  Development by self-digestion: molecular mechanisms and biological functions of autophagy. , 2004, Developmental cell.

[26]  I. Germano,et al.  Role of autophagy in temozolomide-induced cytotoxicity for malignant glioma cells , 2004, Cell Death and Differentiation.

[27]  I. Germano,et al.  Inhibition of DNA repair for sensitizing resistant glioma cells to temozolomide. , 2003, Journal of neurosurgery.

[28]  M. Junier,et al.  PEA‐15 Modulates TNFα Intracellular Signaling in Astrocytes , 2003 .

[29]  Isabelle Callebaut,et al.  The multifunctional protein PEA-15 is involved in the control of apoptosis and cell cycle in astrocytes. , 2003, Biochemical pharmacology.

[30]  G. Condorelli,et al.  Protein Kinase B/Akt Binds and Phosphorylates PED/PEA-15, Stabilizing Its Antiapoptotic Action , 2003, Molecular and Cellular Biology.

[31]  Guido Reifenberger,et al.  Pten signaling in gliomas. , 2002, Neuro-oncology.

[32]  A. Musti,et al.  Multiple Members of the Mitogen-activated Protein Kinase Family Are Necessary for PED/PEA-15 Anti-apoptotic Function* , 2002, The Journal of Biological Chemistry.

[33]  David W. Anderson,et al.  SP600125, an anthrapyrazolone inhibitor of Jun N-terminal kinase , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[34]  E. Formstecher,et al.  PEA-15 mediates cytoplasmic sequestration of ERK MAP kinase. , 2001, Developmental cell.

[35]  E. Liu,et al.  Molecular profiling of transformed and metastatic murine squamous carcinoma cells by differential display and cDNA microarray reveals altered expression of multiple genes related to growth, apoptosis, angiogenesis, and the NF-kappaB signal pathway. , 2001, Cancer research.

[36]  E. Formstecher,et al.  Death effector domain protein PEA-15 potentiates Ras activation of extracellular signal receptor-activated kinase by an adhesion-independent mechanism. , 2000, Molecular biology of the cell.

[37]  M. Gorospe,et al.  c-Jun N-terminal Kinase Is Essential for Growth of Human T98G Glioblastoma Cells* , 2000, The Journal of Biological Chemistry.

[38]  E. Wagner,et al.  Oncogenic transformation by ras and fos is mediated by c-Jun N-terminal phosphorylation , 2000, Oncogene.

[39]  K. Gallo,et al.  Cdc42-induced activation of the mixed-lineage kinase SPRK in vivo. Requirement of the Cdc42/Rac interactive binding motif and changes in phosphorylation. , 2000, The Journal of biological chemistry.

[40]  D. Hanahan,et al.  The Hallmarks of Cancer , 2000, Cell.

[41]  H. Hibshoosh,et al.  Induction of autophagy and inhibition of tumorigenesis by beclin 1 , 1999, Nature.

[42]  J. Girault,et al.  Endothelin Induces a Calcium‐Dependent Phosphorylation of PEA‐15 in Intact Astrocytes: Identification of Ser104 and Ser116 Phosphorylated, Respectively, by Protein Kinase C and Calcium/Calmodulin Kinase II In Vitro , 1998, Journal of neurochemistry.

[43]  Y. Yonekawa,et al.  PTEN (MMAC1) Mutations Are Frequent in Primary Glioblastomas (de novo) but not in Secondary Glioblastomas , 1998, Journal of neuropathology and experimental neurology.

[44]  D. Louis,et al.  PTEN mutations in gliomas and glioneuronal tumors , 1998, Oncogene.

[45]  C. Der,et al.  p120 GAP Modulates Ras Activation of Jun Kinases and Transformation* , 1997, The Journal of Biological Chemistry.

[46]  Michael E. Greenberg,et al.  Opposing Effects of ERK and JNK-p38 MAP Kinases on Apoptosis , 1995, Science.

[47]  D. Grignon,et al.  Loss of heterozygosity of the BRCA1 and other loci on chromosome 17q in human prostate cancer. , 1995, Cancer research.

[48]  Y. Nakamura,et al.  Detailed deletion mapping of chromosome 17q in ovarian and breast cancers: 2-cM region on 17q21.3 often and commonly deleted in tumors. , 1993, Cancer research.

[49]  J. Woodgett,et al.  Phorbol ester-induced amino-terminal phosphorylation of human JUN but not JUNB regulates transcriptional activation. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[50]  M. Karin,et al.  Oncoprotein-mediated signalling cascade stimulates c-Jun activity by phosphorylation of serines 63 and 73 , 1992, Molecular and cellular biology.

[51]  J. Barrett,et al.  Detection of frequent allelic loss on proximal chromosome 17q in sporadic breast carcinoma using microsatellite length polymorphisms. , 1992, Cancer research.

[52]  W. Lowry,et al.  Allele loss from chromosome 17 in ovarian cancer. , 1990, Oncogene.

[53]  Y. Nakamura,et al.  Allele losses on chromosome 17 in human epithelial ovarian carcinoma. , 1990, Oncogene.

[54]  U. K. Laemmli,et al.  A factor preventing the major head protein of bacteriophage T4 from random aggregation. , 1970, Journal of molecular biology.

[55]  Devrim Gozuacik,et al.  Autophagy and cell death. , 2007, Current topics in developmental biology.

[56]  J. Rhim,et al.  C-Jun NH(2)-terminal kinase mediates proliferation and tumor growth of human prostate carcinoma. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[57]  D. Mercola,et al.  Role for c-jun N-terminal kinase in treatment-refractory acute myeloid leukemia (AML): signaling to multidrug-efflux and hyperproliferation , 2002, Leukemia.

[58]  H. Nakshatri,et al.  Regulation of the c-jun gene in p210 BCR-ABL transformed cells corresponds with activity of JNK, the c-jun N-terminal kinase. , 1998, Blood.