Hydrogen Peroxide Activates p70S6k Signaling Pathway*

We investigated a possible role of reactive oxygen species (ROS) in p70S6k activation, which plays an important role in the progression of cells from G0/G1 to S phase of the cell cycle by translational up-regulation of a family of mRNA transcripts that encode for components of the protein synthetic machinery. Treatment of mouse epidermal cell JB6 with H2O2 generated extracellularly by glucose/glucose oxidase led to the activation of p70S6k and p90Rsk and to phosphorylation of p42MAPK/p44MAPK. The activation of p70S6k and p90Rsk was dose-dependent and transient, maximal activities being in extracts treated for 15 and 30 min, respectively. Further characterization of ROS-induced activation of p70S6kusing specific inhibitors for p70S6k signaling pathway, rapamycin, and wortmannin revealed that ROS acted upstream of the rapamycin-sensitive component FRAP/RAFT and wortmannin-sensitive component phosphatidylinositol 3-kinase, because both inhibitors caused the inhibition of ROS-induced p70S6k activity. In addition, Ca2+ chelation also inhibited ROS-induced activation of p70S6k, indicating that Ca2+ is a mediator of p70S6k activation by ROS. However, down-regulation of 12-O-tetradecanoylphorbol-13-acetate (TPA)-responsive protein kinase C (PKC) by chronic pretreatment with TPA or a specific PKC inhibitor Ro-31-8220 did not block the activation of p70S6kby ROS, indicating that the activation of TPA-responsive PKC was not required for stimulation of p70S6k activity by H2O2 in JB6 cells. Exposure of JB6 cells to platelet-derived growth factor or epidermal growth factor led to a rapid increase in H2O2, phosphorylation, and activation of p70S6k, which were antagonized by the pretreatment of catalase. Taken together, the results suggest that ROS act as a messenger in growth factor-induced p70S6k signaling pathway.

[1]  J. Blenis,et al.  p70 S6 Kinase Is Regulated by Protein Kinase Cζ and Participates in a Phosphoinositide 3-Kinase-Regulated Signalling Complex , 1999, Molecular and Cellular Biology.

[2]  J. Avruch,et al.  Atypical protein kinase Clambda binds and regulates p70 S6 kinase. , 1998, The Biochemical journal.

[3]  M. Kasuga,et al.  Requirement for Activation of the Serine-Threonine Kinase Akt (Protein Kinase B) in Insulin Stimulation of Protein Synthesis but Not of Glucose Transport , 1998, Molecular and Cellular Biology.

[4]  J. Turkson,et al.  Role of Tyrosine Kinase Activity of Epidermal Growth Factor Receptor in the Lysophosphatidic Acid-stimulated Mitogen-activated Protein Kinase Pathway* , 1998, The Journal of Biological Chemistry.

[5]  K. Ha,et al.  Phosphatidic Acid-induced Elevation of Intracellular Ca2+ Is Mediated by RhoA and H2O2in Rat-2 Fibroblasts* , 1998, The Journal of Biological Chemistry.

[6]  Tao Yan,et al.  Overexpression of Manganese Superoxide Dismutase Suppresses Tumor Necrosis Factor-induced Apoptosis and Activation of Nuclear Transcription Factor-κB and Activated Protein-1* , 1998, The Journal of Biological Chemistry.

[7]  R. Pearson,et al.  Differential Regulation by Calcium Reveals Distinct Signaling Requirements for the Activation of Akt and p70S6k * , 1998, The Journal of Biological Chemistry.

[8]  M. Andjelkovic,et al.  Phosphorylation and activation of p70s6k by PDK1. , 1998, Science.

[9]  Y Nishizuka,et al.  Activation of protein kinase C by tyrosine phosphorylation in response to H2O2. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[10]  J. Zweier,et al.  Mitogenic Signaling Mediated by Oxidants in Ras-Transformed Fibroblasts , 1997, Science.

[11]  L. Graves,et al.  An Intracellular Calcium Signal Activates p70 but Not p90 Ribosomal S6 Kinase in Liver Epithelial Cells* , 1997, The Journal of Biological Chemistry.

[12]  E. Tekle,et al.  Epidermal Growth Factor (EGF)-induced Generation of Hydrogen Peroxide , 1997, The Journal of Biological Chemistry.

[13]  T. Cruz,et al.  Reactive Oxygen Species Mediate Cytokine Activation of c-Jun NH2-terminal Kinases* , 1996, The Journal of Biological Chemistry.

[14]  M. Andjelkovic,et al.  Activation and phosphorylation of a pleckstrin homology domain containing protein kinase (RAC-PK/PKB) promoted by serum and protein phosphatase inhibitors. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[15]  Qingbo Xu,et al.  Activation of Mitogen-activated Protein Kinase by HO , 1996, The Journal of Biological Chemistry.

[16]  R. Pearson,et al.  The principal target of rapamycin‐induced p70s6k inactivation is a novel phosphorylation site within a conserved hydrophobic domain. , 1995, The EMBO journal.

[17]  V. Ferrans,et al.  Requirement for Generation of H2O2 for Platelet-Derived Growth Factor Signal Transduction , 1995, Science.

[18]  S. Schreiber,et al.  Control of p70 S6 kinase by kinase activity of FRAP in vivo , 1995, Nature.

[19]  R. Pearson,et al.  Rapamycin, Wortmannin, and the Methylxanthine SQ20006 Inactivate p70s6k by Inducing Dephosphorylation of the Same Subset of Sites (*) , 1995, The Journal of Biological Chemistry.

[20]  J. Downward.  A target for Pl(3) kinase , 1995, Nature.

[21]  J. Avruch,et al.  Phosphatidylinositol 3-kinase signals activation of p70 S6 kinase in situ through site-specific p70 phosphorylation. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[22]  T. Cruz,et al.  Involvement of Reactive Oxygen Species in Cytokine and Growth Factor Induction of c-fos Expression in Chondrocytes(*) , 1995, The Journal of Biological Chemistry.

[23]  C. Heldin,et al.  Activation of p70/p85 S6 kinase by a pathway independent of p21fi ras , 1994, Nature.

[24]  K. Nose,et al.  Production of hydrogen peroxide by transforming growth factor-beta 1 and its involvement in induction of egr-1 in mouse osteoblastic cells , 1994, The Journal of cell biology.

[25]  G. Thomas,et al.  p70s6k/p85s6k: mechanism of activation and role in mitogenesis. , 1994, Seminars in cancer biology.

[26]  Andrius Kazlauskas,et al.  PDGF- and insulin-dependent pp70S6k activation mediated by phosphatidylinositol-3-OH kinase , 1994, Nature.

[27]  J. Blenis,et al.  Phosphatidylinositol 3-kinase activation is required for insulin stimulation of pp70 S6 kinase, DNA synthesis, and glucose transporter translocation , 1994, Molecular and cellular biology.

[28]  Stuart L. Schreiber,et al.  A mammalian protein targeted by G1-arresting rapamycin–receptor complex , 1994, Nature.

[29]  G. Thomas,et al.  Rapamycin selectively represses translation of the "polypyrimidine tract" mRNA family. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[30]  A. Fernández,et al.  Nuclear localization of p85s6k: functional requirement for entry into S phase. , 1994, The EMBO journal.

[31]  G. Thomas,et al.  Elongation factor-1 alpha mRNA is selectively translated following mitogenic stimulation. , 1994, The Journal of biological chemistry.

[32]  Z. Kaprielian,et al.  The molecular basis of retinotectal topography , 1994, BioEssays : news and reviews in molecular, cellular and developmental biology.

[33]  H. Lane,et al.  p70s6k function is essential for G1 progression , 1993, Nature.

[34]  J. Avruch,et al.  Rapamycin-induced inhibition of the 70-kilodalton S6 protein kinase. , 1992, Science.

[35]  G. Crabtree,et al.  Rapamycin selectively inhibits interleukin-2 activation of p70 S6 kinase , 1992, Nature.

[36]  G. Crabtree,et al.  Rapamycin-FKBP specifically blocks growth-dependent activation of and signaling by the 70 kd S6 protein kinases , 1992, Cell.

[37]  E. J. Lewis,et al.  The design and biological properties of potent and selective inhibitors of protein kinase C. , 1992, Biochemical Society transactions.

[38]  P. Baeuerle,et al.  Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF‐kappa B transcription factor and HIV‐1. , 1991, The EMBO journal.

[39]  P. Baeuerle,et al.  A role for oxygen radicals as second messengers. , 1991, Trends in cell biology.

[40]  C. Nathan,et al.  Production of large amounts of hydrogen peroxide by human tumor cells. , 1991, Cancer research.

[41]  G. Thomas,et al.  MAP2 kinase and 70K S6 kinase lie on distinct signalling pathways , 1991, Nature.

[42]  J. Avruch,et al.  Molecular structure of a major insulin/mitogen-activated 70-kDa S6 protein kinase. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[43]  M. Siegmann,et al.  Cloning of the mitogen-activated S6 kinase from rat liver reveals an enzyme of the second messenger subfamily. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M. Dale,et al.  The effect of new potent selective inhibitors of protein kinase C on the neutrophil respiratory burst. , 1990, Biochemical and biophysical research communications.

[45]  G. Habermehl,et al.  Human fibroblasts release reactive oxygen species in response to interleukin-1 or tumour necrosis factor-alpha. , 1989, The Biochemical journal.

[46]  D. Alcorta,et al.  Sequence and expression of chicken and mouse rsk: homologs of Xenopus laevis ribosomal S6 kinase , 1989, Molecular and cellular biology.

[47]  P. Cerutti,et al.  Oxidants induce phosphorylation of ribosomal protein S6. , 1988, The Journal of biological chemistry.

[48]  J. Maller,et al.  Insulin-stimulated MAP-2 kinase phosphorylates and activates ribosomal protein S6 kinase II , 1988, Nature.

[49]  G. Thomas,et al.  Differential regulation of S6 phosphorylation by insulin and epidermal growth factor in Swiss mouse 3T3 cells: insulin activation of type 1 phosphatase. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[50]  J. Blenis,et al.  A Xenopus ribosomal protein S6 kinase has two apparent kinase domains that are each similar to distinct protein kinases. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[51]  J. Blenis,et al.  The 70 kDa S6 kinase: regulation of a kinase with multiple roles in mitogenic signalling. , 1995, Current opinion in cell biology.

[52]  S. Ferrari,et al.  S6 phosphorylation and the p70s6k/p85s6k. , 1994, Critical reviews in biochemistry and molecular biology.

[53]  P. Parker,et al.  CHAPTER 8 – PROTEIN KINASE C , 1993 .

[54]  H. Aebi,et al.  Catalase in vitro. , 1984, Methods in enzymology.