Abstract
The extracellular-regulated kinases ERK1 and ERK2 (commonly referred to as ERK1/2) have a crucial role in cardiac hypertrophy. ERK1/2 is activated by mitogen-activated protein kinase kinase-1 (MEK1) and MEK2 (commonly referred to as MEK1/2)-dependent phosphorylation in the TEY motif of the activation loop, but how ERK1/2 is targeted toward specific substrates is not well understood. Here we show that autophosphorylation of ERK1/2 on Thr188 directs ERK1/2 to phosphorylate nuclear targets known to cause cardiac hypertrophy. Thr188 autophosphorylation requires the activation and assembly of the entire Raf-MEK-ERK kinase cascade, phosphorylation of the TEY motif, dimerization of ERK1/2 and binding to G protein βγ subunits released from activated Gq. Thr188 phosphorylation of ERK1/2 was observed in isolated cardiomyocytes induced to undergo hypertrophic growth, in mice upon stimulation of Gq-coupled receptors or after aortic banding and in failing human hearts. Experiments using transgenic mouse models carrying mutations at the Thr188 phosphorylation site of ERK2 suggested a causal relationship to cardiac hypertrophy. We propose that specific phosphorylation events on ERK1/2 integrate differing upstream signals (Raf1-MEK1/2 or G protein–coupled receptor–Gq) to induce cardiac hypertrophy.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Rent or buy this article
Prices vary by article type
from$1.95
to$39.95
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Sugden, P.H. & Clerk, A. Cellular mechanisms of cardiac hypertrophy. J. Mol. Med. 76, 725–746 (1998).
Wettschureck, N. et al. Absence of pressure overload induced myocardial hypertrophy after conditional inactivation of Gαq/Gα11 in cardiomyocytes. Nat. Med. 7, 1236–1240 (2001).
Lips, D.J., deWindt, L.J. & van Kraaij, D.J.W. & Doevendans, P.A. Molecular determinants of myocardial hypertrophy and failure: alternative pathways for beneficial and maladaptive hypertrophy. Eur. Heart J. 24, 883–896 (2003).
Muslin, A.J. Role of Raf proteins in cardiac hypertrophy and cardiomyocyte survival. Trends Cardiovasc. Med. 15, 225–229 (2005).
Glennon, P.E. et al. Depletion of mitogen-activated protein kinase using an antisense oligodeoxynucleotide approach downregulates the phenylephrine-induced hypertrophic response in rat cardiac myocytes. Circ. Res. 78, 954–961 (1996).
Bueno, O.F. et al. The MEK1–ERK1/2 signaling pathway promotes compensated cardiac hypertrophy in transgenic mice. EMBO J. 19, 6341–6350 (2000).
Takeishi, Y. et al. Src and multiple MAP kinase activation in cardiac hypertrophy and congestive heart failure under chronic pressure-overload: comparison with acute mechanical stress. J. Mol. Cell. Cardiol. 33, 1637–1648 (2001).
Haq, S. et al. Differential activation of signal transduction pathways in human hearts with hypertrophy versus advanced heart failure. Circulation 103, 670–677 (2001).
Purcell, N.H. et al. Genetic inhibition of cardiac ERK1/2 promotes stress-induced apoptosis and heart failure but has no effect on hypertrophy in vivo. Proc. Natl. Acad. Sci. USA 104, 14074–14079 (2007).
Rossomando, A.J. et al. Identification of Tyr-185 as the site of tyrosine autophosphorylation of recombinant mitogen-activated protein kinase p42mapk. Proc. Natl. Acad. Sci. USA 89, 5779–5783 (1992).
Seger, R. & Krebs, E.G. The MAPK signalling cascade. FASEB J. 9, 726–735 (1995).
Luttrell, L.M. 'Location, location, location': activation and targeting of MAP kinases by G protein–coupled receptors. J. Mol. Endocrinol. 30, 117–126 (2003).
Caunt, C.J., Finch, A.R., Sedgley, K.R. & McArdle, C.A. Seven-transmembrane receptor signalling and ERK compartmentalization. Trends Endocrinol. Metab. 17, 276–283 (2006).
Torii, S., Nakayama, K., Yamamoto, T. & Nishida, E. Regulatory mechanisms and function of ERK MAP kinases. J. Biochem. 136, 557–561 (2004).
Tohgo, A. et al. The stability of the G protein–coupled receptor–β-arrestin interaction determines the mechanism and functional consequence of ERK activation. J. Biol. Chem. 278, 6258–6267 (2003).
Crespo, P., Xu, N., Simonds, W.F. & Gutkind, J.S. Ras-dependent activation of MAP kinase pathway mediated by G-protein βγ subunits. Nature 369, 418–420 (1994).
Slupsky, J.R. et al. Binding of Gβγ subunits to cRaf1 downregulates G protein-coupled receptor signalling. Curr. Biol. 9, 971–974 (1999).
Faure, M., Voyno-Yasenetskaya, T.A. & Bourne, H.R. cAMP and βγ subunits of heterotrimeric G proteins stimulate the mitogen-activated protein kinase pathway in COS-7 cells. J. Biol. Chem. 269, 7851–7854 (1994).
Schwindinger, W.F. & Robishaw, J.D. Heterotrimeric G protein βγ dimers in growth and differentiation. Oncogene 20, 1653–1660 (2001).
Tachibana, H., Naga Prasad, S.V., Lefkowitz, R.J., Koch, W.J. & Rockman, H.A. Level of β-adrenergic receptor kinase 1 inhibition determines degree of cardiac dysfunction after chronic pressure overload-induced heart failure. Circulation 111, 591–597 (2005).
Li, Z. et al. Effects of two Gβγ-binding proteins—N-terminally truncated phosducin and β-adrenergic receptor kinase C terminus (βARKct)—in heart failure. Gene Ther. 10, 1354–1361 (2003).
Mansour, S.J. et al. Transformation of mammalian cells by constitutively active MAP kinase kinase. Science 265, 966–970 (1994).
Schmitt, J.P. et al. Consequences of pressure overload on sarcomere protein mutation–induced hypertrophic cardiomyopathy. Circulation 108, 1133–1138 (2003).
Thomas, W.G. et al. Adenoviral-directed expression of the type 1A angiotensin receptor promotes cardiomyocyte hypertrophy via transactivation of the epidermal growth factor receptor. Circ. Res. 90, 135–142 (2002).
Brown, J.H. et al. Pathways and roadblocks in muscarinic receptor–mediated growth regulation. Life Sci. 60, 1077–1084 (1997).
Sakata, Y., Hoit, B.D., Liggett, S.B., Walsh, R.A. & Dorn, G.W. II. Decompensation of pressure-overload hypertrophy in Gαq-overexpressing mice. Circulation 97, 1488–1495 (1998).
Akhter, S.A. et al. Targeting the receptor-Gq interface to inhibit in vivo pressure overload myocardial hypertrophy. Science 280, 574–577 (1998).
Rohrbach, S. et al. Neuregulin in cardiac hypertrophy in rats with aortic stenosis. Differential expression of erbB2 and erbB4 receptors. Circulation 100, 407–412 (1999).
Baliga, R.R. et al. NRG-1–induced cardiomyocyte hypertrophy. Role of PI-3-kinase, p70(S6K), and MEK-MAPK-RSK. Am. J. Physiol. 277, H2026–H2037 (1999).
Lorenz, K., Lohse, M.J. & Quitterer, U. Protein kinase C switches the Raf kinase inhibitor from Raf-1 to GRK-2. Nature 426, 574–579 (2003).
Umemori, H. et al. Activation of the G protein Gq/11 through tyrosine phosphorylation of the α subunit. Science 276, 1878–1881 (1997).
Luo, J., Busillo, J.M. & Benovic, J.L. M3 muscarinic acetylcholine receptor–mediated signaling is regulated by distinct mechanisms. Mol. Pharmacol. 74, 338–347 (2008).
Khokhlatchev, A.V. et al. Phosphorylation of the MAP kinase ERK2 promotes its homodimerization and nuclear translocation. Cell 93, 605–615 (1998).
Hartkamp, J., Troppmair, J. & Rapp, U.R. The JNK/SAPK activator mixed lineage kinase 3 (MLK3) transforms NIH 3T3 cells in a MEK-dependent fashion. Cancer. Res. 59, 2195–2202 (1999).
Bogoyevitch, M.A. & Sudgen, P.H. The role of protein kinases in adaptational growth of the heart. Int. J. Biochem. Cell. Biol. 28, 1–12 (1996).
Bueno, O.F. & Molkentin, J.D. Involvement of extracellular signal–regulated kinases 1/2 in cardiac hypertrophy and cell death. Circ. Res. 91, 776–781 (2002).
Zhong, W. et al. Hypertrophic growth in cardiac myocytes is mediated by Myc through a cyclin D2–dependent pathway. EMBO J. 25, 3869–3879 (2006).
Markou, T., Hadzopoulou-Cladarasb, M. & Lazou, A. Phenylephrine induces activation of CREB in adult rat cardiac myocytes through MSK1 and PKA signaling pathways. J. Mol. Cell. Cardiol. 37, 1001–1011 (2004).
Valks, D.M. et al. Phenylephrine promotes phosphorylation of Bad in cardiac myocytes through the extracellular signal–regulated kinases 1/2 and protein kinase A. J. Mol. Cell. Cardiol. 34, 749–763 (2002).
Dorn, G.W. II & Force, T. Protein kinase cascades in regulation of cardiac hypertrophy. J. Clin. Invest. 115, 527–537 (2005).
Garrington, T.P. & Johnson, G.L. Organization and regulation of mitogen-activated protein kinase signaling pathways. Curr. Opin. Cell Biol. 11, 211–218 (1999).
Chang, L. & Karin, M. Mammalian MAP kinase signalling cascades. Nature 410, 37–40 (2001).
Pouysségur, J., Volmat, V. & Lenormand, P. Fidelity and spatio-temporal control in MAP kinase (ERKs) signalling. Biochem. Pharmacol. 64, 755–763 (2002).
Brunet, A. et al. Nuclear translocation of p42/p44 mitogen-activated protein kinase is required for growth factor–induced gene expression and cell cycle entry. EMBO J. 18, 664–674 (1999).
Cobb, M.H. & Goldsmith, E.J. Dimerization in MAP kinase signaling. Trends Biochem. Sci. 25, 7–9 (2000).
Müller, S., Heckman, M. & Lohse, M.J. Specific enhancement of β-adrenergic receptor kinase activity by defined Gβ and Gγ subunits. Proc. Natl. Acad. Sci. USA 90, 10439–10443 (1993).
Frank, M., Thümer, L., Lohse, M.J. & Bünemann, M. G protein activation without subunit dissociation depends on Gαi-specific region. J. Biol. Chem. 280, 24584–24590 (2005).
Acknowledgements
We wish to thank U. Quitterer for stimulating discussions during the early phase of this work; M. Maier-Peuschel, M. Vidal, K. Deiß, D. Calebiro, K. Hadameck, M. Frank, K. von Hayn and A. Zürn for their help; M. Fischer, M. Babl, M. Hoffmann, N. Ziegler and C. Dees for excellent technical assistance; S. Jacobs for help with the immunohistochemistry; and U. Zabel and R. Schreck for helpful discussion. This work was supported by the Deutsche Forschungsgemeinschaft, the Leducq Foundation and the Fonds der Chemischen Industrie.
Author information
Authors and Affiliations
Contributions
K.L. performed all of the experiments and their analyses; J.P.S. performed the aortic banding surgery and provided advice for echocardiography; E.M.S. generated the transgenic mouse lines; and M.J.L. and K.L. conceived the study and wrote the manuscript. E.M.S. and particularly J.P.S. provided critical comments on the manuscript.
Corresponding author
Supplementary information
Supplementary Text and Figures
Supplementary Figs. 1–8 and Supplementary Methods (PDF 762 kb)
Rights and permissions
About this article
Cite this article
Lorenz, K., Schmitt, J., Schmitteckert, E. et al. A new type of ERK1/2 autophosphorylation causes cardiac hypertrophy. Nat Med 15, 75–83 (2009). https://doi.org/10.1038/nm.1893
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/nm.1893