The p53-induced Gene-6 (Proline Oxidase) Mediates Apoptosis through a Calcineurin-dependent Pathway*

Proline oxidase is a p53-induced redox gene that can generate reactive oxygen species (ROS) and mediate apoptosis in tumor cells. We report that proline oxidase is a downstream effector in p53-mediated activation of the calcium/calmodulin-dependent phosphatase calcineurin in lung, renal, colon, and ovarian carcinoma cells. The activation of calcineurin by p53 and proline oxidase was detected by activation of the nuclear factor of activated T cells (NFAT), an established indicator of activated calcineurin. Both proline oxidase- and p53-induced activation of NFAT were sensitive to the calcineurin inhibitors cyclosporin A and FK-506, to scavengers of ROS, and to inhibitors of calcium mobilization. A proline oxidase antisense vector suppressed the ability of p53 to up-regulate proline oxidase, activate calcineurin, and induce apoptosis. Moreover, two renal carcinoma-derived mutant p53 proteins were deficient in inducing proline oxidase expression and in activating calcineurin. Inhibitors of calcineurin and calcium mobilization abolished proline oxidase-mediated apoptosis and reduced p53-induced apoptosis. Treatment of colon and ovarian carcinoma cells with the anticancer genotoxic agent etoposide up-regulated both p53 and proline oxidase, activated calcineurin, and induced apoptosis. The etoposide-mediated activation of calcineurin and induction of apoptosis was markedly suppressed by FK-506 calcineurin inhibitor. We propose that proline oxidase mediates apoptosis through the generation of proline-dependent ROS, which then mobilize calcium and activate calcineurin. The activation of calcineurin-regulated transcription factor pathways by proline oxidase might affect gene expression events important to p53 regulation of cell growth and apoptosis.

[1]  M. Michalak,et al.  Calreticulin, Ca2+, and calcineurin - signaling from the endoplasmic reticulum. , 2004, Molecules and cells.

[2]  Bcl-2–Mediated Drug Resistance Inhibition of Apoptosis by Blocking Nuclear Factor of Activated T Lymphocytes (Nfat)-Induced FAS Ligand Transcription , 1999 .

[3]  A. Rao,et al.  Calcineurin Binds the Transcription Factor NFAT1 and Reversibly Regulates Its Activity (*) , 1996, The Journal of Biological Chemistry.

[4]  L. Oberley,et al.  MnSOD inhibits proline oxidase-induced apoptosis in colorectal cancer cells. , 2005, Carcinogenesis.

[5]  Y. Shen,et al.  p53-dependent apoptosis pathways. , 2001, Advances in cancer research.

[6]  Xinbin Chen,et al.  The ferredoxin reductase gene is regulated by the p53 family and sensitizes cells to oxidative stress-induced apoptosis , 2002, Oncogene.

[7]  K. Kinzler,et al.  A model for p53-induced apoptosis , 1997, Nature.

[8]  Weiya Ma,et al.  Involvement of Nuclear Factor of Activated T Cells Activation in UV Response , 2000, The Journal of Biological Chemistry.

[9]  Masanori Osawa,et al.  The role of calcium-binding proteins in the control of transcription: structure to function. , 2002, BioEssays : news and reviews in molecular, cellular and developmental biology.

[10]  I. Graef,et al.  NFAT signaling in vertebrate development. , 2001, Current opinion in genetics & development.

[11]  W P Bennett,et al.  Molecular epidemiology of human cancer risk: gene–environment interactions and p53 mutation spectrum in human lung cancer , 1999, The Journal of pathology.

[12]  Valerie Horsley,et al.  NFAT: ubiquitous regulator of cell differentiation and adaptation. , 2002, The Journal of cell biology.

[13]  P. Hogan,et al.  Transcription factors of the NFAT family: regulation and function. , 1997, Annual review of immunology.

[14]  Jieyuan Jiang,et al.  Differential regulation of cellular target genes by p53 devoid of the PXXP motifs with impaired apoptotic activity , 1999, Oncogene.

[15]  G. Prendergast,et al.  The polyproline region of p53 is required to activate apoptosis but not growth arrest , 1997, Oncogene.

[16]  V. Castranova,et al.  Hydrogen peroxide mediates activation of nuclear factor of activated T cells (NFAT) by nickel subsulfide. , 2001, Cancer research.

[17]  D. Housman,et al.  p53-dependent apoptosis modulates the cytotoxicity of anticancer agents , 1993, Cell.

[18]  G. Crabtree,et al.  NFAT Signaling Choreographing the Social Lives of Cells , 2002, Cell.

[19]  A. Contente,et al.  Tumor-derived mutations within the DNA-binding domain of p53 that phenotypically resemble the deletion of the proline-rich domain , 2000, Oncogene.

[20]  L. Bracco,et al.  The requirement for the p53 proline‐rich functional domain for mediation of apoptosis is correlated with specific PIG3 gene transactivation and with transcriptional repression , 1998, The EMBO journal.

[21]  P. Schmid,et al.  Expression of p53 during mouse embryogenesis. , 1991, Development.

[22]  Y. Rojanasakul,et al.  Vanadate Induces p53 Transactivation through Hydrogen Peroxide and Causes Apoptosis* , 2000, The Journal of Biological Chemistry.

[23]  T. Gourley,et al.  Cutting Edge: The Class II Transactivator Prevents Activation-Induced Cell Death by Inhibiting Fas Ligand Gene Expression1 , 2001, The Journal of Immunology.

[24]  R. Flavell,et al.  Transcription mediated by NFAT is highly inducible in effector CD4+ T helper 2 (Th2) cells but not in Th1 cells , 1997, Molecular and cellular biology.

[25]  L. Donehower,et al.  Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours , 1992, Nature.

[26]  Y. Mo,et al.  Cytotoxic signalling by inhibitors of DNA topoisomerase II. , 2001, Biochemical Society transactions.

[27]  J. Haddad.  Antioxidant and prooxidant mechanisms in the regulation of redox(y)-sensitive transcription factors. , 2002, Cellular signalling.

[28]  K. Davies,et al.  Calcium and oxidative stress: from cell signaling to cell death. , 2002, Molecular immunology.

[29]  T. Wieloch,et al.  Cyclosporin A, But Not FK 506, Protects Mitochondria and Neurons against Hypoglycemic Damage and Implicates the Mitochondrial Permeability Transition in Cell Death , 1998, The Journal of Neuroscience.

[30]  A. Levine,et al.  Identification of a novel p53 functional domain that is necessary for efficient growth suppression. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[31]  J. Manfredi,et al.  Multiple roles of the tumor suppressor p53 , 2002, Current opinion in oncology.

[32]  F. Shibasaki,et al.  Suppression of signalling through transcription factor NF-AT by interactions between calcineurin and Bcl-2 , 1997, Nature.

[33]  A. Rivera,et al.  Proline Oxidase Induces Apoptosis in Tumor Cells, and Its Expression Is Frequently Absent or Reduced in Renal Carcinomas* , 2003, The Journal of Biological Chemistry.

[34]  C. Klee,et al.  Calcineurin: from structure to function. , 2000, Current topics in cellular regulation.

[35]  C. N. Coleman,et al.  Increases in sequence specific DNA binding by p53 following treatment with chemotherapeutic and DNA damaging agents. , 1993, Cancer research.

[36]  C. Wollheim,et al.  Possible role for calmodulin in insulin release. Studies with trifluoperazine in rat pancreatic islets. , 1980, The Journal of clinical investigation.

[37]  Guido Kroemer,et al.  Mitochondrial control of cell death , 2000, Nature Medicine.

[38]  D. Woods,et al.  Regulation of p53 function. , 2001, Experimental cell research.

[39]  A. Baldwin,et al.  p53 Transactivation of the HIV-1 Long Terminal Repeat Is Blocked by PD 144795, a Calcineurin-Inhibitor with Anti-HIV Properties* , 1998, The Journal of Biological Chemistry.

[40]  K. Davies,et al.  Hamster adapt78 mRNA is a Down syndrome critical region homologue that is inducible by oxidative stress. , 1997, Archives of biochemistry and biophysics.

[41]  J C Reed,et al.  Ca2+-induced apoptosis through calcineurin dephosphorylation of BAD. , 1999, Science.

[42]  Jun O. Liu,et al.  Integration of calcineurin and MEF2 signals by the coactivator p300 during T‐cell apoptosis , 2000, The EMBO journal.

[43]  L. Glimcher,et al.  The transcription factor NFAT4 is involved in the generation and survival of T cells. , 1998, Immunity.

[44]  M. Cummings,et al.  Increased p53 mRNA expression in liver and kidney apoptosis. , 1996, Biochimica et biophysica acta.

[45]  A. Levine,et al.  Surfing the p53 network , 2000, Nature.

[46]  J. H. Kim,et al.  Differential roles of hydrogen peroxide and hydroxyl radical in cisplatin-induced cell death in renal proximal tubular epithelial cells. , 2003, The Journal of laboratory and clinical medicine.

[47]  J. Phang,et al.  Alterations in proline metabolic enzymes with mammalian development. , 1976, Metabolism: clinical and experimental.

[48]  Lin Chen,et al.  Transcriptional regulation by calcium, calcineurin, and NFAT. , 2003, Genes & development.

[49]  J. Viola,et al.  NFATC2 transcription factor regulates cell cycle progression during lymphocyte activation: evidence of its involvement in the control of cyclin gene expression , 2002, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[50]  M. Oren,et al.  Decision making by p53: life, death and cancer , 2003, Cell Death and Differentiation.

[51]  N. Katunuma,et al.  Characteristics of proline oxidase in rat tissues. , 1980, Journal of biochemistry.

[52]  D. Valle,et al.  Proline oxidase, encoded by p53-induced gene-6, catalyzes the generation of proline-dependent reactive oxygen species. , 2001, Cancer research.

[53]  W. El-Deiry.  The role of p53 in chemosensitivity and radiosensitivity , 2003, Oncogene.

[54]  M. Billingsley,et al.  Calcium-dependent interaction of calcineurin with bcl-2 in neuronal tissue , 2003, Neuroscience.

[55]  D. Bandyopadhyay,et al.  Hydroxyl radical is the major causative factor in stress-induced gastric ulceration. , 1997, Free radical biology & medicine.

[56]  V. Rotter,et al.  Introduction: p53 – the first twenty years , 1999, Cellular and Molecular Life Sciences CMLS.

[57]  D. Faller,et al.  Oncogenic Ras Mediates Apoptosis in Response to Protein Kinase C Inhibition through the Generation of Reactive Oxygen Species* , 2000, The Journal of Biological Chemistry.

[58]  M. Torres.  Mitogen-activated protein kinase pathways in redox signaling. , 2003, Frontiers in bioscience : a journal and virtual library.

[59]  P. Jeffrey,et al.  Crystal structure of a p53 tumor suppressor-DNA complex: understanding tumorigenic mutations. , 1994, Science.

[60]  M. Yaffe,et al.  Affinity-driven peptide selection of an NFAT inhibitor more selective than cyclosporin A. , 1999, Science.

[61]  V. Ferrans,et al.  Reactive oxygen species are downstream mediators of p53-dependent apoptosis. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[62]  T. Russo,et al.  Gene regulation by reactive oxygen species. , 1997, Current topics in cellular regulation.

[63]  J. Hescheler,et al.  Reactive Oxygen Species as Intracellular Messengers During Cell Growth and Differentiation , 2001, Cellular Physiology and Biochemistry.