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:

Phosphorylation-dependent activation of the Ras-GRF/CDC25Mm exchange factor by muscarinic receptors and G-protein βγ subunits

Abstract

MUSCARINIC receptors activate Ras through a pathway distinct1,2 from that mediated through translocation of the exchange factor mSos1 by receptor tyrosine kinases3,4. Here we report that muscarinic receptors can activate another Ras exchange factor, CDC25Mm, or p140Ras-GRF (refs 5,6). In NIH-3T3 cells expressing subtype 1 human muscarinic receptors (hml), the agonist carbachol selectively increased the specific activity and phosphorylation state of epitope-tagged Ras-GRF. This stimulation was reversed by protein phosphatase 1 (PP1), and prevented by transducin α-subunits. Carbachol treatment of neonatal rat brain explants increased Ras exchange factor activity and the phosphorylation state of endogenous Ras-GRF. In COS-7 cells, cotransfection of hml or hm2 receptors with Ras-GRF conferred carbachol-dependent increases in exchange-factor activity, whereas cotransfection with G-protein βγ subunits caused a constitutive activation that was sensitive to PP1. These results demonstrate a G-protein-coupled mechanism for Ras activation, mediated by p140Ras-GRF.

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

Access options

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

  1. Winitz, S. et al. J. biol. Chem. 268, 19196–19199 (1993).

    CAS  Google Scholar 

  2. Mattingly, R. R., Sorisky, A., Brann, M. R. & Macara, I. G. Molec. cell. Biol. 14, 7943–7952 (1994).

    Article  CAS  Google Scholar 

  3. Egan, S. E. et al. Nature 363, 45–51 (1993).

    Article  ADS  CAS  Google Scholar 

  4. Buday, L. & Downward, J. Cell 73, 611–620 (1993).

    Article  CAS  Google Scholar 

  5. Martegani, E. et al. EMBO J. 11, 2151–2157 (1992).

    Article  CAS  Google Scholar 

  6. Shou, C., Farnsworth, C. L., Neel, B. G. & Feig, L. A. Nature 358, 351–354 (1992).

    Article  ADS  CAS  Google Scholar 

  7. Boguski, M. S. & McCormick, F. Nature 366, 643–654 (1993).

    Article  ADS  CAS  Google Scholar 

  8. van Corven, E. J., Hordijk, P. L., Medema, R. H., Bos, J. L. & Moolenaar, W. H. Proc. natn. Acad. Sci. U.S.A. 90, 1257–1261 (1993).

    Article  ADS  CAS  Google Scholar 

  9. van Biesen, T. et al. Nature 376, 781–784 (1995).

    Article  ADS  CAS  Google Scholar 

  10. Downward, J., Graves, J. D., Warne, P. H., Rayter, S. & Cantrell, D. A. Nature 346, 719–723 (1990).

    Article  ADS  CAS  Google Scholar 

  11. Farnsworth, C. L. et al. Nature 376, 524–527 (1995).

    Article  ADS  CAS  Google Scholar 

  12. Guerrero, C. et al. Oncogene 12, 1097–1107 (1996).

    CAS  PubMed  Google Scholar 

  13. Shou, C., Wurmser, A., Ling, K., Barbacid, M. & Feig, L. A. Oncogene 10, 1887–1893 (1995).

    CAS  Google Scholar 

  14. Cen, H., Papageorge, A. G., Vass, W. C., Zhang, K. & Lowy, D. R. Molec. cell. Biol. 13, 7718–7724 (1993).

    Article  CAS  Google Scholar 

  15. Hawes, B. E., van Biesen, T., Koch, W. J., Luttrell, L. M. & Lefkowitz, R. J. J. biol. Chem. 270, 17148–17153 (1995).

    Article  CAS  Google Scholar 

  16. Wolfman, A. & Macara, I. G. Science 248, 67–69 (1990).

    Article  ADS  CAS  Google Scholar 

  17. Gutkind, J. S., Novotny, E. A., Brann, M. R. & Robbins, K. C. Proc. natn. Acad. Sci. U.S.A. 88, 4703–4707 (1991).

    Article  ADS  CAS  Google Scholar 

  18. Koch, W. J., Hawes, B. E., Allen, L. F. & Lefkowitz, R. J. Proc. natn. Acad. Sci. U.S.A. 91, 12706–12710 (1994).

    Article  ADS  CAS  Google Scholar 

  19. Crespo, P., Xu, N., Simmonds, W. F. & Gutkind, J. S. Nature 369, 418–420 (1994).

    Article  ADS  CAS  Google Scholar 

  20. Faure, M., Voyno-Yasenetskaya, T. A. & Bourne, H. R. J. biol. Chem. 269, 7851–7854 (1994).

    CAS  Google Scholar 

  21. Porfiri, E. & McCormick, F. J. biol. Chem. 271, 5871–5877 (1996).

    Article  CAS  Google Scholar 

  22. LaMorte, V. J. et al. J. biol. Chem. 269, 13490–13496 (1994).

    CAS  PubMed  Google Scholar 

  23. Hanley, M. R. Nature 340, 97 (1989).

    Article  ADS  CAS  Google Scholar 

  24. Ramirez, M. T., Post, G. R., Sulakhe, P. V. & Brown, J. H. J. biol. Chem. 270, 8446–8451 (1995).

    Article  CAS  Google Scholar 

  25. Thorburn, A. et al. J. biol. Chem. 268, 2244–2249 (1993).

    CAS  PubMed  Google Scholar 

  26. Posada, J. & Cooper, J. A. Science 255, 212–215 (1992).

    Article  ADS  CAS  Google Scholar 

  27. Mattingly, R. R., Wasilenko, W. J., Woodring, P. J. & Garrsion, J. C. J. biol. Chem. 267, 7470–7477 (1992).

    CAS  PubMed  Google Scholar 

  28. Linnemann, D. in A Dissection and Tissue Culture Manual of the Nervous System (ed. Shahar, A. et al.) 75–76 (Alan R. Liss, New York, 1989).

    Google Scholar 

  29. Graber, S. G., Figler, R. A., Kalman-Maltese, V. K., Robishaw, J. D. & Garrison, J. C. J. biol. Chem. 267, 13123–13126 (1992).

    CAS  PubMed  Google Scholar 

  30. Brondyk, W. H., McKiernan, C. J., Burstein, E. S. & Macara, I. G. J. biol. Chem. 268, 9410–9415 (1993).

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Mattingly, R., Macara, I. Phosphorylation-dependent activation of the Ras-GRF/CDC25Mm exchange factor by muscarinic receptors and G-protein βγ subunits. Nature 382, 268–272 (1996). https://doi.org/10.1038/382268a0

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/382268a0

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

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