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.

Growth factor-induced MAPK network topology shapes Erk response determining PC-12 cell fate

An Addendum to this article was published on 01 May 2007

Abstract

The mitogen-activated protein kinase (MAPK) network is a conserved signalling module that regulates cell fate by transducing a myriad of growth-factor signals1. The ability of this network to coordinate and process a variety of inputs from different growth-factor receptors into specific biological responses is, however, still not understood. We investigated how the MAPK network brings about signal specificity in PC-12 cells, a model for neuronal differentiation2. Reverse engineering by modular-response analysis3 uncovered topological differences in the MAPK core network dependent on whether cells were activated with epidermal or neuronal growth factor (EGF or NGF). On EGF stimulation, the network exhibited negative feedback only, whereas a positive feedback was apparent on NGF stimulation. The latter allows for bi-stable Erk activation dynamics, which were indeed observed. By rewiring these regulatory feedbacks, we were able to reverse the specific cell responses to EGF and NGF. These results show that growth factor context determines the topology of the MAPK signalling network and that the resulting dynamics govern cell fate.

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

Relevant articles

Open Access articles citing this article.

Access options

Buy article

Get time limited or full article access on ReadCube.

$32.00

All prices are NET prices.

Figure 1: MRA reveals differences in MAPK-network topology.
Figure 2: Dose-response profiles of Erk1/2 activation.
Figure 3: Irreversibility in MAPK network activation on NGF stimulation.
Figure 4: Rewiring MAPK-network topology redirects phenotypic response of PC-12 cells.

References

  1. O'Neill, E. & Kolch, W. Conferring specificity on the ubiquitous Raf/MEK signalling pathway. Br. J. Cancer 90, 283–288 (2004).

    Article  CAS  Google Scholar 

  2. Greene, L. A. & Tischler, A. S. Establishment of a noradrenergic clonal line of rat adrenal pheochromocytoma cells which respond to nerve growth factor. Proc. Natl Acad. Sci. USA 73, 2424–2428 (1976).

    Article  CAS  Google Scholar 

  3. Kholodenko, B. N. et al. Untangling the wires: a strategy to trace functional interactions in signaling and gene networks. Proc. Natl Acad. Sci. USA 99, 12841–12846 (2002).

    Article  CAS  Google Scholar 

  4. Marshall, C. J. Specificity of receptor tyrosine kinase signaling: transient versus sustained extracellular signal-regulated kinase activation. Cell 80, 179–185 (1995).

    Article  CAS  Google Scholar 

  5. Murphy, L. O., Smith, S., Chen, R. H., Fingar, D. C. & Blenis, J. Molecular interpretation of ERK signal duration by immediate early gene products. Nature Cell Biol. 4, 556–564 (2002).

    Article  CAS  Google Scholar 

  6. Schoeberl, B., Eichler-Jonsson, C., Gilles, E. D. & Muller, G. Computational modeling of the dynamics of the MAP kinase cascade activated by surface and internalized EGF receptors. Nature Biotechnol. 20, 370–375 (2002).

    Article  Google Scholar 

  7. Kao, S., Jaiswal, R. K., Kolch, W. & Landreth, G. E. Identification of the mechanisms regulating the differential activation of the mapk cascade by epidermal growth factor and nerve growth factor in PC12 cells. J. Biol. Chem. 276, 18169–18177 (2001).

    Article  CAS  Google Scholar 

  8. Sasagawa, S., Ozaki, Y., Fujita, K. & Kuroda, S. Prediction and validation of the distinct dynamics of transient and sustained ERK activation. Nature Cell Biol. 7, 365–373 (2005).

    Article  CAS  Google Scholar 

  9. Diaz, B. et al. Phosphorylation of Raf-1 serine 338-serine 339 is an essential regulatory event for Ras-dependent activation and biological signaling. Mol. Cell Biol. 17, 4509–4516 (1997).

    Article  CAS  Google Scholar 

  10. Anderson, N. G., Maller, J. L., Tonks, N. K. & Sturgill, T. W. Requirement for integration of signals from two distinct phosphorylation pathways for activation of MAP kinase. Nature 343, 651–653 (1990).

    Article  CAS  Google Scholar 

  11. Alessi, D. R. et al. Identification of the sites in MAP kinase kinase-1 phosphorylated by p74raf-1. EMBO J. 13, 1610–1619 (1994).

    Article  CAS  Google Scholar 

  12. Andrec, M., Kholodenko, B. N., Levy, R. M. & Sontag, E. Inference of signaling and gene regulatory networks by steady-state perturbation experiments: structure and accuracy. J. Theor. Biol. 232, 427–441 (2005).

    Article  CAS  Google Scholar 

  13. Eblen, S. T. et al. Mitogen-activated protein kinase feedback phosphorylation regulates MEK1 complex formation and activation during cellular adhesion. Mol. Cell Biol. 24, 2308–2317 (2004).

    Article  CAS  Google Scholar 

  14. Frost, J. A. et al. Cross-cascade activation of ERKs and ternary complex factors by Rho family proteins. EMBO J. 16, 6426–6438 (1997).

    Article  CAS  Google Scholar 

  15. Buday, L., Warne, P. H. & Downward, J. Downregulation of the Ras activation pathway by MAP kinase phosphorylation of Sos. Oncogene 11, 1327–1331 (1995).

    CAS  PubMed  Google Scholar 

  16. Ferrell, J. E., Jr. & Machleder, E. M. The biochemical basis of an all-or-none cell fate switch in Xenopus oocytes. Science 280, 895–898 (1998).

    Article  CAS  Google Scholar 

  17. Ferrell, J. E., Jr. Building a cellular switch: more lessons from a good egg. Bioessays 21, 866–870 (1999).

    Article  Google Scholar 

  18. Kholodenko, B. N. Cell-signalling dynamics in time and space. Nature Rev. Mol. Cell. Biol. 7, 165–176 (2006).

    Article  CAS  Google Scholar 

  19. Bhalla, U. S., Ram, P. T. & Iyengar, R. MAP kinase phosphatase as a locus of flexibility in a mitogen-activated protein kinase signaling network. Science 297, 1018–1023 (2002).

    Article  CAS  Google Scholar 

  20. Corbit, K. C. et al. Activation of Raf-1 signaling by protein kinase C through a mechanism involving Raf kinase inhibitory protein. J. Biol. Chem. 278, 13061–13068 (2003).

    Article  CAS  Google Scholar 

  21. 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).

    Article  CAS  Google Scholar 

  22. Yeung, K. et al. Suppression of Raf-1 kinase activity and MAP kinase signalling by RKIP. Nature 401, 173–177 (1999).

    Article  CAS  Google Scholar 

  23. Balan, V. et al. Identification of novel in vivo Raf-1 phosphorylation sites mediating positive feedback Raf-1 regulation by extracellular signal-regulated kinase. Mol. Biol. Cell 17, 1141–1153 (2006).

    Article  CAS  Google Scholar 

  24. Krutzik, P. O. & Nolan, G. P. Intracellular phospho-protein staining techniques for flow cytometry: monitoring single cell signaling events. Cytometry 55, 61–70 (2003).

    Article  Google Scholar 

Download references

Acknowledgements

We are thankful to G. Nolan for FACS protocols and to A. Riddell (Flow Cytometry Core Facility, EMBL) for technical assistance. We also thank P. Cohen, A. Squire, P. Beltrao and A. Kinkhabwala for very helpful discussions. S. Santos is supported by an 'E-STAR' fellowship funded by the ECs FP6 Marie Curie Host fellowship for Early Stage Research Training under contract number MEST-CT-2004-504640.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Philippe I. H. Bastiaens.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary figures S1, S2, S3 and S4 (PDF 1624 kb)

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Santos, S., Verveer, P. & Bastiaens, P. Growth factor-induced MAPK network topology shapes Erk response determining PC-12 cell fate. Nat Cell Biol 9, 324–330 (2007). https://doi.org/10.1038/ncb1543

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ncb1543

This article is cited by

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