Anisomycin-activated protein kinases p45 and p55 but not mitogen-activated protein kinases ERK-1 and -2 are implicated in the induction of c-fos and c-jun

Independent of its ability to block translation, anisomycin intrinsically initiates intracellular signals and immediate-early gene induction [L. C. Mahadevan and D. R. Edwards, Nature (London) 349:747-749, 1991]. Here, we characterize further its action as a potent, selective signalling agonist. In-gel kinase assays show that epidermal growth factor (EGF) transiently activates five kinases: the mitogen-activated protein (MAP) kinases ERK-1 and -2, and three others, p45, p55, and p80. Anisomycin, at inhibitory and subinhibitory concentrations, does not activate ERK-1 and -2 but elicits strong sustained activation of p45 and p55, which are unique in being serine kinases whose detection is enhanced with poly-Glu/Tyr or poly-Glu/Phe copolymerized in these gels. Translational arrest using emetine or puromycin does not activate p45 and p55 but does prolong EGF-stimulated ERK-1 and -2 activation. Rapamycin, which blocks anisomycin-stimulated p70/85S6k activation without affecting nuclear responses, has no effect on p45 or p55 kinase. p45 and p55 are activable by okadaic acid or UV irradiation, and both kinases phosphorylate the c-Jun NH2-terminal peptide 1-79, putatively placing them within c-Jun NH2-terminal kinase/stress-activated protein kinase (JNK/SAPK) subfamily of MAP kinases. Thus, the EGF- and anisomycin-activated kinases p45 and p55 are strongly implicated in signalling to c-fos and c-jun, whereas the MAP kinases ERK-1 and -2 are not essential for this process.

[1]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[2]  M J Barratt,et al.  Mitogen-stimulated phosphorylation of histone H3 is targeted to a small hyperacetylation-sensitive fraction. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[3]  J. Woodgett,et al.  The stress-activated protein kinase subfamily of c-Jun kinases , 1994, Nature.

[4]  G. Johnson,et al.  Sequential protein kinase reactions controlling cell growth and differentiation. , 1994, Current opinion in cell biology.

[5]  M. Karin,et al.  JNK1: A protein kinase stimulated by UV light and Ha-Ras that binds and phosphorylates the c-Jun activation domain , 1994, Cell.

[6]  R. Treisman.  Ternary complex factors: growth factor regulated transcriptional activators. , 1994, Current opinion in genetics & development.

[7]  C. Marshall,et al.  MAP kinase kinase kinase, MAP kinase kinase and MAP kinase. , 1994, Current opinion in genetics & development.

[8]  L. Mahadevan,et al.  Anisomycin and rapamycin define an area upstream of p70/85S6k containing a bifurcation to histone H3-HMG-like protein phosphorylation and c-fos-c-jun induction , 1994 .

[9]  Hong Sun,et al.  MKP-1 (3CH134), an immediate early gene product, is a dual specificity phosphatase that dephosphorylates MAP kinase in vivo , 1993, Cell.

[10]  M. Karin,et al.  Identification of an oncoprotein- and UV-responsive protein kinase that binds and potentiates the c-Jun activation domain. , 1993, Genes & development.

[11]  A. Brunet,et al.  Growth factors induce nuclear translocation of MAP kinases (p42mapk and p44mapk) but not of their activator MAP kinase kinase (p45mapkk) in fibroblasts , 1993, The Journal of cell biology.

[12]  G. Pfeifer,et al.  In vivo protein-DNA interactions at the c-jun promoter: preformed complexes mediate the UV response , 1993, Molecular and cellular biology.

[13]  J. Blenis,et al.  Signal transduction via the MAP kinases: proceed at your own RSK. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[14]  R. Hipskind,et al.  c‐fos transcriptional activation and repression correlate temporally with the phosphorylation status of TCF. , 1993, The EMBO journal.

[15]  R. Treisman,et al.  The SRF accessory protein Elk-1 contains a growth factor-regulated transcriptional activation domain , 1993, Cell.

[16]  E. Nishida,et al.  The MAP kinase cascade is essential for diverse signal transduction pathways. , 1993, Trends in biochemical sciences.

[17]  S. Winawer,et al.  Signal transduction through extracellular signal-regulated kinase-like pp57 blocked in differentiated cells having low protein kinase C beta activity. , 1993, The Journal of biological chemistry.

[18]  E. Nishida,et al.  cDNA cloning of MAP kinase kinase reveals kinase cascade pathways in yeasts to vertebrates. , 1993, The EMBO journal.

[19]  J. Woodgett,et al.  Co-purification of mitogen-activated protein kinases with phorbol ester-induced c-Jun kinase activity in U937 leukaemic cells. , 1993, Oncogene.

[20]  R. Davis,et al.  Mitogen-activated protein kinase stimulation by a tyrosine kinase-negative epidermal growth factor receptor. , 1993, The Journal of biological chemistry.

[21]  S. Keyse,et al.  Oxidative stress and heat shock induce a human gene encoding a protein-tyrosine phosphatase , 1992, Nature.

[22]  D. Meek,et al.  Nuclear protein phosphorylation and growth control. , 1992, The Biochemical journal.

[23]  S. Ferrari,et al.  Activation of p70s6k is associated with phosphorylation of four clustered sites displaying Ser/Thr-Pro motifs. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[24]  A. Sharrocks,et al.  Phosphorylation of transcription factor p62TCF by MAP kinase stimulates ternary complex formation at c-fos promoter , 1992, Nature.

[25]  R. Campos-González,et al.  Tyrosine phosphorylation of mitogen-activated protein kinase in cells with tyrosine kinase-negative epidermal growth factor receptors. , 1992, The Journal of biological chemistry.

[26]  D. Edwards,et al.  Protein synthesis inhibitors differentially superinduce c‐fos and c‐jun by three distinct mechanisms: lack of evidence for labile repressors. , 1992, The EMBO journal.

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

[28]  S. Pelech,et al.  Mitogen-activated protein kinases: versatile transducers for cell signaling. , 1992, Trends in biochemical sciences.

[29]  L. Heasley,et al.  MAP kinase is constitutively activated in gip2 and src transformed rat 1a fibroblasts. , 1992, The Journal of biological chemistry.

[30]  J. Blenis,et al.  Nuclear localization and regulation of erk- and rsk-encoded protein kinases , 1992, Molecular and cellular biology.

[31]  J. Avruch,et al.  An array of insulin-activated, proline-directed serine/threonine protein kinases phosphorylate the p70 S6 kinase. , 1992, The Journal of biological chemistry.

[32]  G. Thomas.  MAP kinase by any other name smells just as sweet , 1992, Cell.

[33]  James R. Woodgett,et al.  Phosphorylation of c-jun mediated by MAP kinases , 1991, Nature.

[34]  H. Kawasaki,et al.  Xenopus M phase MAP kinase: isolation of its cDNA and activation by MPF. , 1991, The EMBO journal.

[35]  J. Avruch,et al.  pp54 microtubule-associated protein-2 kinase requires both tyrosine and serine/threonine phosphorylation for activity. , 1991, The Journal of biological chemistry.

[36]  J. Blenis,et al.  Distinct mechanisms for the activation of the RSK kinases/MAP2 kinase/pp90rsk and pp70-S6 kinase signaling systems are indicated by inhibition of protein synthesis. , 1991, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[37]  Anthony C. Willis,et al.  Rapid histone H3 phosphorylation in response to growth factors, phorbol esters, okadaic acid, and protein synthesis inhibitors , 1991, Cell.

[38]  Nancy Y. Ip,et al.  ERKs: A family of protein-serine/threonine kinases that are activated and tyrosine phosphorylated in response to insulin and NGF , 1991, Cell.

[39]  D. Burns,et al.  Lipid activation of protein kinase C. , 1991, The Journal of biological chemistry.

[40]  L. Mahadevan,et al.  Signalling and superinduction , 1991, Nature.

[41]  L. Cantley,et al.  Oncogenes and signal transduction , 1991, Cell.

[42]  P. Majerus,et al.  Recent insights in phosphatidylinositol signaling , 1990, Cell.

[43]  J. Ferrell,et al.  Identification of a 42-kilodalton phosphotyrosyl protein as a serine(threonine) protein kinase by renaturation , 1990, Molecular and cellular biology.

[44]  A E Nel,et al.  Stimulation of MAP-2 kinase activity in T lymphocytes by anti-CD3 or anti-Ti monoclonal antibody is partially dependent on protein kinase C. , 1990, Journal of immunology.

[45]  M. Rosenfeld,et al.  Hormonally inducible phosphorylation of a nuclear pool of ribosomal protein S6. , 1990, The Journal of biological chemistry.

[46]  A. Saltiel,et al.  Nerve growth factor stimulates a protein kinase in PC-12 cells that phosphorylates microtubule-associated protein-2. , 1990, The Journal of biological chemistry.

[47]  P. Cohen,et al.  Okadaic acid: a new probe for the study of cellular regulation. , 1990, Trends in biochemical sciences.

[48]  M. Sheng,et al.  The inner core of the serum response element mediates both the rapid induction and subsequent repression of c-fos transcription following serum stimulation. , 1990, Genes & development.

[49]  F. C. Lucibello,et al.  Trans-repression of the mouse c-fos promoter: A novel mechanism of fos-mediated trans-regulation , 1989, Cell.

[50]  M. Siegmann,et al.  A stimulated S6 kinase from rat liver: identity with the mitogen activated S6 kinase of 3T3 cells. , 1989, The EMBO journal.

[51]  I. Kameshita,et al.  A sensitive method for detection of calmodulin-dependent protein kinase II activity in sodium dodecyl sulfate-polyacrylamide gel. , 1989, Analytical biochemistry.

[52]  Richard Treisman,et al.  Removal of poly(A) and consequent degradation of c-fos mRNA facilitated by 3′ AU-rich sequences , 1988, Nature.

[53]  M. Yaniv,et al.  Transcriptional activation of c-jun during the G0/G1 transition in mouse fibroblasts , 1988, Nature.

[54]  R. Bravo,et al.  Complexity of the early genetic response to growth factors in mouse fibroblasts , 1988, Molecular and cellular biology.

[55]  E. Nishida,et al.  Activation of a Ca2+-inhibitable protein kinase that phosphorylates microtubule-associated protein 2 in vitro by growth factors, phorbol esters, and serum in quiescent cultured human fibroblasts. , 1988, The Journal of biological chemistry.

[56]  I. Verma,et al.  Modification of fos proteins: phosphorylation of c-fos, but not v-fos, is stimulated by 12-tetradecanoyl-phorbol-13-acetate and serum , 1987, Molecular and cellular biology.

[57]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[58]  M. Greenberg,et al.  Effect of protein synthesis inhibitors on growth factor activation of c-fos, c-myc, and actin gene transcription , 1986, Molecular and cellular biology.

[59]  T. Curran,et al.  FBJ murine osteosarcoma virus: identification and molecular cloning of biologically active proviral DNA , 1982, Journal of virology.

[60]  A. Riggs,et al.  Genomic Sequencing , 2010 .

[61]  A. Nebreda,et al.  Inactivation of MAP kinases. , 1994, Trends in biochemical sciences.

[62]  E. Racker.  Use of synthetic amino acid polymers for assay of protein-tyrosine and protein-serine kinases. , 1991, Methods in enzymology.

[63]  H. Herschman.  Primary response genes induced by growth factors and tumor promoters. , 1991, Annual review of biochemistry.

[64]  D. Marshak,et al.  Synthetic peptide substrates for casein kinase II. , 1991, Methods in enzymology.