Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

An agonist-induced conformational change in the growth hormone receptor determines the choice of signalling pathway

Nature Cell Biology volume 10, pages 740–747 (2008)Cite this article

An Erratum to this article was published on 01 July 2008

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.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: A conformational change in the extracellular domain of the GH receptor related to receptor activation.
Figure 2: Tyr phosphorylation of JAK2 and GHR and STAT5b activation are normal with F'G'ΔGHR in FDC-P1 cells, but ERK signalling through Ras and proliferation are impaired.
Figure 3: Evidence that GH-dependent ERK1/2 activation in FDC-P1 cells expressing GHR is dependent on an SFK (Lyn) constitutively associated with the GHR and can be prevented by PLCγ inhibition.
Figure 4: GH can activate SFK and ERK in JAK2-deficient cells and in mice harbouring an inactive GHR, whereas Lyn binds to the proximal region of GHR in the absence of JAK2.
Figure 5: Perturbing mutations within the linker region above the TMD alter the alignment of the TMDs according to modelling, and alter the ratio of STAT5 to ERK signalling in reporter assays.

Similar content being viewed by others

References

  1. 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).

    Article  CAS  Google Scholar 

  2. Cunningham, B. C. & Wells, J. A. Comparison of a structural and a functional epitope. J. Mol. Biol. 234, 554–563 (1993).

    Article  CAS  Google Scholar 

  3. 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).

    Article  CAS  Google Scholar 

  4. 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).

    Google Scholar 

  5. Harding, P. A. et al. GH and a GH antagonist promote GH receptor dimerization and internalisation. J. Biol. Chem. 271, 6708–6712 (1996).

    Article  CAS  Google Scholar 

  6. 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).

    CAS  PubMed  Google Scholar 

  7. 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).

    Article  CAS  Google Scholar 

  8. 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).

    Article  CAS  Google Scholar 

  9. 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).

    CAS  PubMed  Google Scholar 

  10. Argetsinger, L. S. et al. Identification of JAK2 as a growth hormone receptor-associated tyrosine kinase. Cell 74, 237–244 (1993).

    Article  CAS  Google Scholar 

  11. 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).

    CAS  PubMed  Google Scholar 

  12. Bivona, T. G. et al. Phospholipase C-γ activates Ras on the golgi apparatus by means of RasGRP1. Nature 424, 694–698 (2003)

    Article  CAS  Google Scholar 

  13. 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).

    Article  CAS  Google Scholar 

  14. Kohlhuber, F. et al. A JAK1/JAK2 chimera can sustain α and γ interferon responses. Mol. Cell Biol. 17, 695–706 (1997).

    Article  CAS  Google Scholar 

  15. 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).

    Article  CAS  Google Scholar 

  16. Syed, R. S. et al. Efficiency of signalling through cytokine receptors depends critically on orientation. Nature 395, 511–516 (1998).

    Article  CAS  Google Scholar 

  17. 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).

    CAS  PubMed  Google Scholar 

  18. 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).

    Article  CAS  Google Scholar 

  19. 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).

    Article  CAS  Google Scholar 

  20. 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).

    Article  CAS  Google Scholar 

  21. Clayton, P. E. et al. Signal transduction defects in growth hormone insensitivity. Acta Paediatr. Suppl. 88, 174–178 (1999).

    Article  CAS  Google Scholar 

  22. 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).

    Article  CAS  Google Scholar 

  23. 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).

    Article  CAS  Google Scholar 

  24. 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).

    Article  CAS  Google Scholar 

  25. 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).

    Article  CAS  Google Scholar 

  26. 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).

    Article  CAS  Google Scholar 

  27. 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).

    CAS  PubMed  Google Scholar 

  28. 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).

    CAS  PubMed  Google Scholar 

  29. 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).

    CAS  PubMed  Google Scholar 

  30. 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).

    Article  CAS  Google Scholar 

  31. 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).

    Article  CAS  Google Scholar 

  32. Ptitsyn, O. B. & Finkelstein, A. V. Theory of protein secondary structure and algorithm of its prediction. Biopolymers 22, 15–25 (1983).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by National Health and Medical Research Council (NHMRC, Australia). We thank Liz Holliday for her excellent technical assistance.

Author information

Author notes

  1. Scott W. Rowlinson

    Present address: Current address: Eli Lilly and Company, Lilly Corporate Center, Drop Code 0444, Indianapolis, Indiana, 46285, USA.,

  2. Hideo Yoshizato

    Present address: Current address: Department of Materials Science and Engineering, Nagoya Institue of Technology, Gokiso-cho, Showa-ku, Nagoya 466-8555, Japan.,

Authors and Affiliations

  1. Institute for Molecular Bioscience and School of Biomedical Sciences, University of Queensland, St Lucia, 4072, Australia

    Scott W. Rowlinson, Hideo Yoshizato, Johanna L. Barclay, Andrew J. Brooks, Stuart N. Behncken, Linda M. Kerr, Kirstin Millard, Kathryn Palethorpe, Katherine Nielsen, Jodie Clyde-Smith, John F. Hancock & Michael J. Waters

Authors
  1. Scott W. Rowlinson
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hideo Yoshizato
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Johanna L. Barclay
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Andrew J. Brooks
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stuart N. Behncken
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Linda M. Kerr
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kirstin Millard
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kathryn Palethorpe
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Katherine Nielsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Jodie Clyde-Smith
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. John F. Hancock
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Michael J. Waters
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael J. Waters.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Figures S1, S2, Supplementary Table 1 and Supplementary Discussion (PDF 704 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1737

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing