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Nur factors are critical for proopiomelanocortin (POMC) induction by CRH in corticotrophs, but the pathways linking CRH to Nur are unknown. In this study we show that in AtT-20 corticotrophs CRH and cAMP induce Nur77 and Nurr1 expression and transcription at the NurRE site by protein kinase A (PKA) and calcium-dependent and -independent mechanisms. Calcium pathways depend on calmodulin kinase II (CAMKII) activity, and calcium-independent pathways are accounted for in part by MAPK activation (Rap1/B-Raf/MAPK-ERK kinase/ERK1/2), demonstrated by the use of molecular and pharmacological tools. ATT-20 corticotrophs express B-Raf, as do other cells in which cAMP stimulates MAPK. CRH/cAMP stimulated ERK2 activity and increased transcriptional activity of a Gal4-Elk1 protein, which was blocked by overexpression of dominant negative mutants and kinase inhibitors and stimulated by expression of B-Raf. The MAPK kinase inhibitors did not affect Nur77 and Nurr1 mRNA induction but blocked CRH or cAMP-stimulated Nur transcriptional activity. Moreover, MAPK stimulated phosphorylation and transactivation of Nur77. The functional impact of these pathways was confirmed at the POMC promoter. In conclusion, in AtT-20 corticotrophs the CRH/cAMP signaling that leads to Nur77/Nurr1 mRNA induction and transcriptional activation, and thus POMC expression, is dependent on protein kinase A and involves calcium/calmodulin kinase II (Nur induction/activation) and MAPK calcium-dependent and -independent (Nur phosphorylation-activation) pathways.


Documento: Artículo
Título:Activation and induction of Nur77/Nurr1 in corticotrophs by CRH/cAMP: Involvement of calcium, protein kinase a, and MAPK pathways
Autor:Kovalovsky, D.; Refojo, D.; Liberman, A.C.; Hochbaum, D.; Pereda, M.P.; Coso, O.A.; Stalla, G.K.; Holsboer, F.; Arzt, E.
Filiación:Laboratorio de Fisiología y Biología Molecular, Departamento de Biología, Universidad de Buenos Aires, 1428 Buenos Aires, Argentina
Max Planck Institute of Psychiatry, 80804 Munich, Germany
Laboratorio de Fisiología y Biología Molecular, Facultad de Ciencias Exactas y Naturales, Ciudad Universitaria (1428), Buenos Aires, Argentina
Palabras clave:corticosteroid receptor; cyclic AMP dependent protein kinase; mitogen activated protein kinase; proopiomelanocortin; protein kinase (calcium,calmodulin); calcium; calcium channel blocking agent; cell receptor; corticotropin releasing factor; cyclic AMP; DNA binding protein; Elk1 protein, mouse; nifedipine; nuclear receptor Nur77; Nurr1 nuclear receptor; oncoprotein; steroid receptor; transcription factor; transcription factor Elk 1; animal cell; article; cell type; corticotropin release; enzyme activation; enzyme induction; gene overexpression; hypothalamus hypophysis adrenal system; nonhuman; point mutation; priority journal; protein phosphorylation; stimulus response; stress; transcription regulation; animal; cell culture; cytology; DNA responsive element; drug effect; genetic transcription; genetics; hypophysis; metabolism; mouse; mutation; phosphorylation; promoter region; signal transduction; Animalia; Animals; Calcium; Calcium Channel Blockers; Cells, Cultured; Corticotropin-Releasing Hormone; Cyclic AMP; Cyclic AMP-Dependent Protein Kinases; DNA-Binding Proteins; ets-Domain Protein Elk-1; MAP Kinase Signaling System; Mice; Mutation; Nifedipine; Phosphorylation; Pituitary Gland; Pro-Opiomelanocortin; Promoter Regions (Genetics); Proto-Oncogene Proteins; Receptors, Cytoplasmic and Nuclear; Receptors, Steroid; Response Elements; Signal Transduction; Transcription Factors; Transcription, Genetic
Página de inicio:1638
Página de fin:1651
Título revista:Molecular Endocrinology
Título revista abreviado:Mol. Endocrinol.
CAS:cyclic AMP dependent protein kinase; mitogen activated protein kinase, 142243-02-5; proopiomelanocortin, 66796-54-1; calcium, 7440-70-2, 14092-94-5; corticotropin releasing factor, 9015-71-8, 178359-01-8, 79804-71-0, 86297-72-5, 86784-80-7; cyclic AMP, 60-92-4; nifedipine, 21829-25-4; Calcium, 7440-70-2; Calcium Channel Blockers; Corticotropin-Releasing Hormone, 9015-71-8; Cyclic AMP, 60-92-4; Cyclic AMP-Dependent Protein Kinases, EC, 7440-70-2; DNA-Binding Proteins; Elk1 protein, mouse; ets-Domain Protein Elk-1; Nifedipine, 21829-25-4, 9015-71-8; Nurr1 nuclear receptor; orphan nuclear receptor NGFI-B, 121479-42-3; Pro-Opiomelanocortin, 66796-54-1; Proto-Oncogene Proteins; Receptors, Cytoplasmic and Nuclear; Receptors, Steroid; Transcription Factors


  • Lundblad, J.R., Roberts, J.L., Regulation of proopiomelanocortin gene expression in pituitary (1988) Endocr Rev, 9, pp. 135-158
  • Levin, N., Blum, M., Roberts, J.L., Modulation of basal and corticotropin-releasing factor-stimulated proopiomelanocortin gene expression by vasopressin in rat anterior pituitary (1989) Endocrinology, 125, pp. 2957-2966
  • Boutillier, A.L., Sassone-Corsi, P., Loeffler, J.P., The protooncogene c-fos is induced by corticotropin-releasing factor and stimulates proopiomelanocortin gene transcription in pituitary cells (1991) Mol Endocrinol, 5, pp. 1301-1310
  • Boutillier, A.L., Monnier, D., Lorang, D., Lundblad, J.R., Roberts, J.L., Loeffler, J.P., Corticotropin-releasing hormone stimulates proopiomelanocortin transcription by cFos-dependent and -independent pathways: Characterization of an AP1 site in exon 1 (1995) Mol Endocrinol, 9, pp. 745-755
  • Usui, T., Nakai, Y., Tsukada, T., Fukata, J., Nakaishi, S., Naitoh, Y., Imura, H., Cyclic AMP-responsive region of the human proopiomelanocortin (POMC) gene (1989) Mol Cell Endocrinol, 62, pp. 141-146
  • Schecterson, L.C., Mcknight, G.S., Role of cyclic adenosine 3′,5′-monophosphate-dependent protein kinase in hormone-stimulated β-endorphin secretion in AtT20 cells (1991) Mol Endocrinol, 5, pp. 170-178
  • Timpl, P., Spanagel, R., Sillaber, I., Kresse, A., Reul, J.M., Stalla, G.K., Blanquet, V., Wurst, W., Impaired stress response anal reduced anxiety in mice lacking a functional corticotropin-releasing hormone receptor (1998) Nat Genet, 19, pp. 162-166
  • Luini, A., Lewis, D., Guild, S., Corda, D., Axelrod, J., Hormone secretagogues increase cytosolic calcium by increasing cAMP in corticotropin-secreting cells (1985) Proc Natl Acad Sci USA, 82, pp. 8034-8038
  • Litvin, Y., PasMantier, R., Fleischer, N., Erlichman, J., Hormonal activation of the cAMP-dependent protein kinases in AtT20 cells (1984) J Biol Chem, 259, pp. 10296-10302
  • Stalla, G.K., Stalla, J., Huber, M., Loeffler, J.P., Hollt, V., Von Werder, K., Muller, O.A., Ketoconazole inhibits corticotropic cell function in vitro (1988) Endocrinology, 122, pp. 618-623
  • Lee, A.K., Tse, A., Mechanism undedying corticotropin-releasing hormone (CRH) triggered cytosolic Ca rise in identified rat corticotrophs (1997) J Physiol, 504, pp. 367-378
  • Clark, T.P., Kemppainen, R.J., Glucocorticoids do not affect intracellular calcium transients in corticotrophs: Evidence supporting an effect distal to calcium influx (1994) Neuroendocrinology, 60, pp. 273-282
  • Von Dreden, G., Loeffler, J.P., Grimm, C., Hollt, V., Influence of calcium ions on proopiomelanocortin mRNA levels in anterior clonal pituitary cells (1988) Neuroendocrinology, 47, pp. 32-37
  • Philips, A., Lesage, S., Gingras, R., Maira, M.H., Gauthier, Y., Hugo, P., Drouin, J., Novel dimeric Nur77 signaling mechanism in endocrine and lymphoid cells (1997) Mol Cell Biol, 17, pp. 5946-5951
  • Maira, M., Martens, C., Philips, A., Drouin, J., Heterodimedzation between members of the Nur subfamily of orphan nuclear receptors as a novel mechanism for gene activation (1999) Mol Cell Biol, 19, pp. 7549-7557
  • Murphy, E.P., Coneely, O.M., Neuroendocrine regulation of the hypothalamic pituitary adrenal axis by the nurr1/nur77 subfamily of nuclear receptors (1997) Mol Endocrinol, 11, pp. 39-47
  • Emmark, E., Gustafsson, J.A., Orphan nuclear receptors-the first eight years (1996) Mol Endocrinol, 10, pp. 1293-1307
  • Wilson, T.E., Mouw, A.R., Weaver, C.A., Milbrandt, J., Parker, K.L., The orphan nuclear receptor NGFI-B regulates expression of the gene encoding steroid-21-hydroxylase (1993) Mol Cell Biol, 13, pp. 861-868
  • Philips, A., Maira, M., Mullick, A., Chamberland, M., Lesage, S., Hugo, P., Drouin, J., Antagonism between Nur77 and glucocorticoid receptor for control of transcription (1997) Mol Cell Biol, 17, pp. 5952-5959
  • Altschuler, D., Lapetina, E.G., Mutational analysis of the cAMP-dependent protein kinase-mediated phosphorylation site of Rap1b (1993) J Biol Chem, 268, pp. 7527-7531
  • Rooij, J., Zwartkruis, F.J.T., Verheijen, M.H.G., Cool, R.H., Nijman, S.M.B., Wittinghofer, A., Bos, J.L., Epac is a Rap1 guanine-nucleotide-exchange factor directly activated by cyclic AMP (1998) Nature, 396, pp. 474-477
  • Cook, S.J., Rubinfeld, B., Albert, I., McCormick, F., RapV12 antagonizes Ras-dependent activation of ERK1 and ERK2 by LPA and EGF in rat-1 fibroblasts (1993) EMBO J, 12, pp. 3475-3485
  • Li, W., Whaley, C.D., Mondino, A., Müller, D.L., Blocked signal transduction to the ERK ancl JNK protein kinases in anergic CD4+ T cells (1996) Science, 271, pp. 1272-1276
  • Fields, P.E., Gajewski, T.F., Fitch, F.W., Blocked Ras activation in anergic CD4+ T cells (1996) Science, 271, pp. 1276-1278
  • Bos, J.L., All in the family? New insights and questions regarding interconnectivity of Ras, Rap1 and Ral (1998) EMBO J, 17, pp. 6776-6782
  • Cook, S.J., McCormick, F., Inhibition by cAMP of Ras-dependent activation of Raf (1993) Science, 262, pp. 1069-1072
  • Wu, J., Dent, P., Jelinek, T., Wolfman, A., Weber, M.J., Sturgill, T.W., Inhibition of the EGF-activated MAP kinase signaling pathway by adenosine 3′,5′-monophosphate (1993) Science, 262, pp. 1065-1069
  • Vossler, M.R., Yao, H., York, R.D., Pan, M.G., Rim, C.S., Stork, P.J.S., cAMP activates MAP kinase and Elk-1 through a B-Raf-and Rap1-dependent pathway (1997) Cell, 89, pp. 73-82
  • Dugan, L.L., Kim, J.S., Zhang, Y., Bart, R.D., Sun, Y., Holtzman, D.M., Gutmann, D.H., Differential effects of cAMP in neurons and astrocytes. Role of B-raf (1999) J Biol Chem, 274, pp. 25842-25848
  • Young, S.W., Dickens, M., Tavare, J.M., Differentiation of PC12 cells in response to a cAMP analogue is accompanied by sustained activation of mitogen-activated protein kinase: Comparison with the effects of insulin, growth factors and phorbol esters (1994) FEBS Lett, 338, pp. 212-216
  • York, R.D., Yao, H., Dillon, T., Ellig, C.L., Eckert, S.P., McCleskey, E.W., Stork, P.J.S., Rap1 mediates sustained MAP kinase activation induced by nerve growth factor (1998) Nature, 392, pp. 622-626
  • Qiu, W., Zhuang, S., Von Lintig, F.C., Boss, G.R., Pilz, R.B., Cell type-specific regulation of B-raf kinase by cAMP and 14-3-3 proteins (2000) J Biol Chem, 275, pp. 31921-31929
  • Kievit, P., Lauten, J.D., Maurer, R.A., Analysis of the role of the mitogen-activated protein kinase in mediating cyclic-adenosine 3′,5′-monophosphate effects on prolactin promoter activity (2001) Mol Endocrinol, 15, pp. 614-624
  • Richards, J.S., New signaling pathways for hormones and cyclic adenosine 3′,5′-monophosphate action in endocrine cells (2001) Mol Endocrinol, 15, pp. 209-218
  • Woronicz, J.D., Lina, A., Calnan, B.J., Szychowski, S., Cheng, L., Winoto, A., Regulation of the Nur77 orphan steroid receptor in activation-induced apoptosis (1995) Mol Cell Biol, 15, pp. 6364-6376
  • Youn, H.D., Sun, L., Prywes, R., Liu, J.O., Apoptosis of T cells mediated by Ca-induced release of the transcription factor MEF2 (1999) Science, 286, pp. 790-793
  • Blaeser, F., Ho, N., Prywes, R., Chatila, T.A., Ca(2+)-dependent gene expression mediated by MEF2 transcription factors (2000) J Biol Chem, 275, pp. 197-209
  • Castillo, S.O., Xiao, Q., Lyu, M.S., Kozak, C.A., Nikodem, V.M., Organization, sequence, chromosomal localization, and promoter identification of the mouse orphan nuclear receptor Nurr1 gene (1997) Genomics, 41 (2), pp. 250-257
  • Antaraki, A., Ang, K.L., Antoni, F.A., Involvement of calyculin A inhibitable protein phosphatases in the cyclic AM P signal transduction pathway of mouse corticotroph tumor (AtT20) cells (1997) Br J Pharmacol, 121, pp. 991-999
  • Erlichman, J., Litvin, Y., Fleischer, N., Immunological and molecular characterization of cAMP-dependent protein kinases in AtT-20 cells (1984) J Biol Chem, 259, pp. 10289-10295
  • Tsygankova, O.M., Saavedra, A., Rebhun, J.F., Quilliam, L.A., Meinkoth, J.L., Coordinated regulation of Rap1 and thyroid differentiation by cyclic AMP and protein kinase A (2001) Mol Cell Biol, 21, pp. 1921-1929
  • Grewal, S.S., Horgan, A.M., York, R.D., Withers, G.S., Bankers, G.A., Stork, P.J., Neuronal calcium activates Rap1 and B-Raf signaling pathway via the cyclic adenosine monophosphate-dependent protein kinase (2000) J Biol Chem, 275, pp. 3722-3728
  • Egea, J., Espinet, C., Soler, R.M., Peiro, S., Rocamora, N., Comella, J., Nerve growth factor activation of the extracellular signal-regulated kinase pathway is modulated by Ca+ and calmodulin (2000) Mol Cell Biol, 20, pp. 1931-1946
  • Grewal, S.S., Fass, D.M., Yao, H., Ellig, C.L., Goodman, R.H., Stork, P.J.S., Calcium and cAMP signals differentially regurate cAMP-responsive element-binding protein function via a Rap1-extracellular signal-regulated kinase pathway (2000) J Biol Chem, 275, pp. 34433-34441
  • Seternes, O.M., Johansen, B., Moens, U., A dominant role for the Raf-MEK pathway in forskolin, 12-O-tetradecanoyl-phorbol acetate, and platelet-derived growth factor-induced CREB (cAMP-responsive element-binding protein) activation, uncoupled from serine 133 phosphorylation in NIH 3T3 cells (1999) Mol Endocrinol, 13, pp. 1071-1083
  • Arzt, E., Stelzer, G., Renner, U., Lange, M., Müller, O.A., Stalla, G.K., Interleukin-2 and interleukin-2 receptor expression in human corticotrophic adenoma and murine pituitary cell cultures (1992) J Clin Invest, 90, pp. 1944-1951
  • Ohkura, N., Hosono, T., Maruyama, K., Tsukada, T., Yamaguchi, K., An isoform of Nurr1 as a negative inhibitor of the NGFI-B family signaling (1999) Biochim Biophys Acta, 1444, pp. 69-79
  • Liu, B., Mortrud, M., Low, M.J., DNA elements with AT-rich core sequences direct pituitary cell-specific expression of the pro-opiomelanocortin gene in transgenic mice (1995) Biochem J, 312, pp. 827-832
  • Peterson, S.N., Trabalzini, L., Brtva, T.R., Fischer, T., Altschuler, D.L., Martelli, P., Lapetina, E.G., White, G.C., Identification of a novel RalGDS-related protein as a candidate effector for Ras and Rap1 (1996) J Biol Chem, 271, pp. 29903-29908
  • McKean, D.J., Bell, M., Huntoon, C., Rastogi, S., Norstrand, M.V., Podzorski, R., Nilson, A., Paya, C., IL-1 receptor and TCR signals synergize to activate NF-kB mediated gene transcription (1995) Int Immunol, 7, pp. 9-20
  • Rammes, G., Steckler, T., Lresse, A., Schutz, G., Zieglgansberger, W., Lutz, B., Synaptic plasticity in the basolateral amygdala in transgenic mice expressing dominant-negative cAMP response element-binding protein (CREB) in forebrain (2000) Eur J Neurosci, 12, pp. 2534-2546
  • Clegg, C.H., Abrahamsern, M.S., Degen, J.L., Morris, D.R., McKnight, G.S., Cyclic AMP-dependent protein kinase controls basal gene activity and steroidogenesis in Y1 adrenal tumor cells (1992) Biochemistry, 31, pp. 3720-3726
  • Coso, O.A., Chiariello, M., Yu, J.C., Teramoto, H., Crespo, P., Xu, N., Miki, T., Gutkind, J.S., The small GTP-binding proteins Rac1 and Cdc42 regulate the activity of the JNK/SAPK signaling pathway (1995) Cell, 81, pp. 1137-1146
  • Arzt, E., Sauer, J., Pollmacher, T., Labeur, M., Holboer, F., Reul, J.M.H.M., Stalla, G.K., Glucocorticoids suppress interleukin-1 receptor antagonist synthesis following induction by endotoxin (1994) Endocrinology, 134, pp. 672-677
  • Costas, M., Trapp, T., Paez Pereda, M., Sauer, J., Rupprecht, R., Nahmod, V.E., Reul, J.M.H.M., Arzt, E., Molecular and functional evidence for in vitro cytokine enhancement of human and murine target cell sensitivity to grucocorticoids (1996) J Clin Invest, 98, pp. 1409-1416
  • Crespo, P., Xu, N., Simonds, W.F., Gutkind, J.S., Ras-dependent activation of MAPK pathway mediated by G-protein βγ subunits (1994) Nature, 369, pp. 418-420


---------- APA ----------
Kovalovsky, D., Refojo, D., Liberman, A.C., Hochbaum, D., Pereda, M.P., Coso, O.A., Stalla, G.K.,..., Arzt, E. (2002) . Activation and induction of Nur77/Nurr1 in corticotrophs by CRH/cAMP: Involvement of calcium, protein kinase a, and MAPK pathways. Molecular Endocrinology, 16(7), 1638-1651.
---------- CHICAGO ----------
Kovalovsky, D., Refojo, D., Liberman, A.C., Hochbaum, D., Pereda, M.P., Coso, O.A., et al. "Activation and induction of Nur77/Nurr1 in corticotrophs by CRH/cAMP: Involvement of calcium, protein kinase a, and MAPK pathways" . Molecular Endocrinology 16, no. 7 (2002) : 1638-1651.
---------- MLA ----------
Kovalovsky, D., Refojo, D., Liberman, A.C., Hochbaum, D., Pereda, M.P., Coso, O.A., et al. "Activation and induction of Nur77/Nurr1 in corticotrophs by CRH/cAMP: Involvement of calcium, protein kinase a, and MAPK pathways" . Molecular Endocrinology, vol. 16, no. 7, 2002, pp. 1638-1651.
---------- VANCOUVER ----------
Kovalovsky, D., Refojo, D., Liberman, A.C., Hochbaum, D., Pereda, M.P., Coso, O.A., et al. Activation and induction of Nur77/Nurr1 in corticotrophs by CRH/cAMP: Involvement of calcium, protein kinase a, and MAPK pathways. Mol. Endocrinol. 2002;16(7):1638-1651.