Abstract
The growth and metabolic actions of growth hormone (GH) are believed to be mediated through the GH receptor (GHR) by JAK2 activation. The GHR exists as a constitutive homodimer, with signal transduction by ligand-induced realignment of receptor subunits1. Based on the crystal structures2,3, we identify a conformational change in the F'G' loop of the lower cytokine module, which results from binding of hGH but not G120R hGH antagonist. Mutations disabling this conformational change cause impairment of ERK but not JAK2 and STAT5 activation by the GHR in FDC-P1 cells. This results from the use of two associated tyrosine kinases by the GHR, with JAK2 activating STAT5, and Lyn activating ERK1/2. We provide evidence that Lyn signals through phospholipase Cγ, leading to activation of Ras. Accordingly, mice with mutations in the JAK2 association motif respond to GH with activation of hepatic Src and ERK1/2, but not JAK2/STAT5. We suggest that F'G' loop movement alters the signalling choice between JAK2 and a Src family kinase by regulating TMD realignment. Our findings could explain debilitated ERK but not STAT5 signalling in some GH-resistant dwarfs and suggest pathway-specific cytokine agonists.
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References
Brown, R. J. et al. Model for growth hormone receptor activation based on subunit rotation within a receptor dimer. Nature Struct. Mol. Biol. 12, 814–821 (2005).
Cunningham, B. C. & Wells, J. A. Comparison of a structural and a functional epitope. J. Mol. Biol. 234, 554–563 (1993).
De Vos, A. M., Ultsch, M. & Kossiakoff, A. A. Human growth hormone and extracellular domain of its receptor: Crystal structure of the complex. Science 255, 306–312 (1992).
Smit, L., Meyer, D., Argetsinger, L., Schwartz, J. & Carter-Su, C. Molecular events in GH-receptor interaction and signalling. In Handbook of Physiology, Section 7: The Endocrine System. (ed Kostyo, J. L.) 445–480 (Oxford Univ. Press, NY, 1999).
Harding, P. A. et al. GH and a GH antagonist promote GH receptor dimerization and internalisation. J. Biol. Chem. 271, 6708–6712 (1996).
Ross, R. J. et al. Binding and functional studies with the growth hormone receptor antagonist, B2036-PEG (pegvisomant), reveal effects of pegylation and evidence that it binds to a receptor dimer. J. Clin. Endocrinol. Metab. 86, 1716–1723 (2001).
Kopchick, J. J., Parkinson, C., Stevens, E. C. & Trainer, P. J. Growth hormone receptor antagonists: discovery, development, and use in patients with acromegaly. Endocrinol. Rev. 23, 623–646 (2002).
Clackson, T., Ultsch, M. H., Wells, J. A. & de Vos, A. M. Structural and functional analysis of the 1:1 growth hormone:receptor complex reveals the molecular basis for receptor affinity. J. Mol. Biol. 277, 1111–1128 (1998).
Walker, J. L. et al. A novel mutation affecting the interdomain link region of the growth hormone receptor in a Vietnamese girl, and response to long-term treatment with recombinant human insulin-like growth factor-I and luteinizing hormone-releasing hormone analogue. J. Clin. Endocrinol. Metab. 83, 2554–2561 (1998).
Argetsinger, L. S. et al. Identification of JAK2 as a growth hormone receptor-associated tyrosine kinase. Cell 74, 237–244 (1993).
Campbell, G. S., Pang, L., Miyasaka, T., Saltiel, A. R. & Carter-Su, C. Stimulation by growth hormone of MAP kinase activity in 3T3-F442A fibroblasts. J. Biol. Chem. 267, 6074–6080 (1992).
Bivona, T. G. et al. Phospholipase C-γ activates Ras on the golgi apparatus by means of RasGRP1. Nature 424, 694–698 (2003)
Zhu, T., Ling, L. & Lobie, P. E. Identification of a JAK2-independent pathway regulating growth hormone (GH)-stimulated p44/42 mitogen-activated protein kinase activity. GH activation of Ral and phospholipase D is Src-dependent. J. Biol. Chem. 277, 45592–45603 (2002).
Kohlhuber, F. et al. A JAK1/JAK2 chimera can sustain α and γ interferon responses. Mol. Cell Biol. 17, 695–706 (1997).
Livnah, O. et al. An antagonist-peptide EPO receptor complex suggests that receptor dimerization is not sufficient for activation. Nature Struct. Biol. 5, 993–1004 (1998).
Syed, R. S. et al. Efficiency of signalling through cytokine receptors depends critically on orientation. Nature 395, 511–516 (1998).
Baumgartner, J. W., Wells, C. A., Chen, C. M. & Waters, M. J. The role of the WSXWS equivalent motif in growth hormone receptor function. J. Biol. Chem. 269, 29094–29101 (1994).
Chen, C., Brinkworth, R. & Waters, M. J. The role of receptor dimerization domain residues in growth hormone signalling. J. Biol. Chem. 272, 5133–5140 (1997).
Gent, J., Van Den Eijnden, M., Van Kerkhof, P. & Strous, G. J. Dimerization and signal transduction of the growth hormone receptor. Mol. Endocrinol. 17, 967–975 (2003).
Seubert, N. et al. Active and inactive orientations of the transmembrane and cytosolic domains of the erythropoietin receptor dimer. Mol. Cell 12, 1239–1250 (2003).
Clayton, P. E. et al. Signal transduction defects in growth hormone insensitivity. Acta Paediatr. Suppl. 88, 174–178 (1999).
Lewis, M. D. et al. A novel dysfunctional GH variant (Ile179Met) exhibits a decreased ability to activate the ERK pathway. J. Clin. Endocrinol. Metab. 89, 1068–1075 (2004).
Rowlinson, S. W. et al. Activation of chimeric and full-length growth hormone receptors by growth hormone receptor monoclonal antibodies. A specific conformational change may be required for full-length receptor signalling. J. Biol. Chem. 273, 5307–5314 (1998).
Behncken, S. N. et al. Aspartate 171 is the major primate-specific determinant of human growth hormone. Engineering porcine growth hormone to activate the human receptor. J. Biol. Chem. 272, 27077–27083 (1997).
Rowland J. E. et al. In vivo analysis of growth hormone receptor signalling domains and their associated transcripts. Mol. Cell Biol. 25, 66–77 (2005).
Wan Y., Zheng Y. Z., Harris J. M., Brown R. & Waters M. J. Epitope map for a growth hormone receptor agonist monoclonal antibody, MAb 263. Mol. Endocrinol. 17, 2240–2250 (2003).
Colosi, P., Wong, K., Leong, S. R. & Wood, W. I. Mutational analysis of the intracellular domain of the human growth hormone receptor. J. Biol. Chem. 268, 12617–12623 (1993).
Clarkson, R. W. E. et al. Early responses of trans-activating factors to growth hormone in preadipocytes: differential regulation of CCAAT enhancer-binding protein-β (C/EBP β) and C/EBP δ. Mol. Endocrinol. 9, 108–121 (1995).
Porfiri, E., Evans, T., Chardin, P. & Hancock J. F. Prenylation of Ras proteins is required for efficient hSOS1-promoted guanine nucleotide exchange. J. Biol. Chem. 269, 22672–22677 (1994).
Clarkson, R. W. E., Shang, C. A., Levitt, L. K., Howard, T. & Waters, M. J. Ternary complex factors Elk-1 and Sap-1a mediate growth hormone-induced transcription of egr-1 (early growth response factor-1) in 3T3-F442A preadipocytes. Mol. Endocrinol. 13, 619–631 (1999).
Daniel, N., Waters, M. J., Bignon, C. & Djiane, J. Involvement of a subset of tyrosine kinases and phosphatases in regulation of the β-lactoglobulin gene promoter by prolactin. Mol. Cell. Endocrinol. 118, 25–35 (1996).
Ptitsyn, O. B. & Finkelstein, A. V. Theory of protein secondary structure and algorithm of its prediction. Biopolymers 22, 15–25 (1983).
Acknowledgements
This work was supported by National Health and Medical Research Council (NHMRC, Australia). We thank Liz Holliday for her excellent technical assistance.
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Rowlinson, S., Yoshizato, H., Barclay, J. et al. An agonist-induced conformational change in the growth hormone receptor determines the choice of signalling pathway. Nat Cell Biol 10, 740–747 (2008). https://doi.org/10.1038/ncb1737
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DOI: https://doi.org/10.1038/ncb1737
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