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Interaction of a G-protein β-subunit with a conserved sequence in Ste20/PAK family protein kinases

Nature volume 391, pages 191–195 (1998)Cite this article

Abstract

Serine/threonine protein kinases of the Ste20/PAK family have been implicated in the signalling from heterotrimeric G proteins to mitogen-activated protein (MAP) kinase cascades1,2. In the yeast Saccharomyces cerevisiae, Ste20 is involved in transmitting the mating-pheromone signal from the βγ-subunits (encoded by the STE4 and STE18 genes, respectively) of a heterotrimeric G protein to a downstream MAP kinase cascade1. We have identified a binding site for the G-protein β-subunit (Gβ) in the non-catalytic carboxy-terminal regions of Ste20 and its mammalian homologues, the p21-activated protein kinases (PAKs). Association of Gβ with this site in Ste20 was regulated by binding of pheromone to the receptor. Mutations in Gβ and Ste20 that prevented this association blocked activation of the MAP kinase cascade. Considering the high degree of structural and functional conservation of Ste20/PAK family members and G-protein subunits, our results provide a possible model for a role of these kinases in Gβγ-mediated signal transduction in organisms ranging from yeast to mammals.

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Figure 1: Association of Ste4 with Ste20 and Ste5.
Figure 2: In vitro -Gβ binding assays.
Figure 3: In vitro -Gβ binding assays.
Figure 4: Mutational analysis of the association of Ste4 with Ste20.
Figure 5: Mutational analysis of the association of Ste4 with Ste20.
Figure 6

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References

  1. Leberer, E., Thomas, D. Y. & Whiteway, M. Pheromone signalling and polarized morphogenesis in yeast. Curr. Opin. Genet. Dev. 7, 59–66 (1997).

    Article  CAS  Google Scholar 

  2. Sells, M. A. & Chernoff, J. Emerging from the Pak: the p21-activated protein kinase family. Trends Cell Biol. 7, 162–167 (1997).

    Article  CAS  Google Scholar 

  3. Peter, M., Neiman, A. M., Park, H.-O., Lohuizen, L. v. & Herskowitz, I. Functional analysis of the interaction between the small GTP binding protein Cdc42 and the Ste20 protein kinase in yeast. EMBO J. 15, 7046–7059 (1996).

    Article  CAS  Google Scholar 

  4. Leberer, E. et al. Functional characterization of the Cdc42p binding domain of yeast Ste20p protein kinase. EMBO J. 16, 83–97 (1997).

    Article  CAS  Google Scholar 

  5. Chang, F. & Herskowitz, I. Phosphorylation of FAR1 in response to α-factor: a possible requirement for cell-cycle arrest. Mol. Biol. Cell 3, 445–450 (1992).

    Article  CAS  Google Scholar 

  6. Gartner, A., Nasmyth, K. & Ammerer, G. Signal transduction in Saccharomyces cerevisiae requires tyrosine and threonine phosphorylation of FUS3 and KSS1. Genes Dev. 6, 1280–1292 (1992).

    Article  CAS  Google Scholar 

  7. Wu, C. et al. Activation of myosin-I by members of the Ste20p protein kinase family. J. Biol. Chem. 271, 31787–31790 (1996).

    Article  CAS  Google Scholar 

  8. Whiteway, M. S. et al. Association of the yeast pheromone response G protein βγ subunits with the MAP kinase scaffold Ste5p. Science 269, 1572–1575 (1995).

    Article  ADS  CAS  Google Scholar 

  9. Bagrodia, S., Taylor, S. J., Creasy, C. L., Chernoff, J. & Cerione, R. A. Identification of a mouse p21Cdc42/Rac activated kinase. J. Biol. Chem. 270, 22731–22737 (1995).

    Article  CAS  Google Scholar 

  10. Cvrckova, F., Devirgilio, C., Manser, E., Pringle, J. R. & Nasmyth, K. Ste20-like protein kinases are required for normal localization of cell growth and for cytokinesis in budding yeast. Genes Dev. 9, 1817–1830 (1995).

    Article  CAS  Google Scholar 

  11. Leberer, E., Dignard, D., Hougan, L., Thomas, D. Y. & Whiteway, M. Dominant-negative mutants of a yeast G-protein β subunit identify two functional regions involved in pheromone signaling. EMBO J. 13, 4805–4813 (1992).

    Article  Google Scholar 

  12. Wall, M. A. et al. The structure of the G protein heterotrimer Gi α1 β1 γ2. Cell 83, 1047–1058 (1995).

    Article  CAS  Google Scholar 

  13. Sondek, J., Bohm, A., Lambright, D. G., Hamm, H. E. & Sigler, P. B. Crystal structure of a G-protein βγ dimer at 2.1A resolution. Nature 379, 369–374 (1996).

    Article  ADS  CAS  Google Scholar 

  14. Wu, C. L., Whiteway, M., Thomas, D. Y. & Leberer, E. Molecular characterization of Ste20p, a potential mitogen-activated protein or extracellular signal-regulated kinase kinase (MEK) kinase kinase from Saccharomyces cerevisiae. J. Biol. Chem. 270, 15984–15992 (1995).

    Article  CAS  Google Scholar 

  15. Leberer, E. et al. Cloning of Saccharomyces cerevisiae STE5 as a suppressor of a Ste20 protein kinase mutant: structural and functional similarity of Ste5 to Far1. Mol. Gen. Genet. 241, 241–254 (1993).

    CAS  PubMed  Google Scholar 

  16. Sikorski, R. S. & Hieter, P. Asystem of shuttle vectors and yeast host strains designed for efficient manipulation of DNA in Saccharomyces cerevisiae. Genetics 122, 19–27 (1989).

    CAS  PubMed  PubMed Central  Google Scholar 

  17. Leberer, E., Dignard, D., Harcus, D., Thomas, D. Y. & Whiteway, M. The protein kinase homologue Ste20p is required to link the yeast pheromone response G-protein βγ subunits to downstream signalling components. EMBO J. 11, 4815–4824 (1992).

    Article  CAS  Google Scholar 

  18. Kunkel, T. D., Roberts, J. D. & Zakour, R. A. Rapid and efficient site-specific mutagenesis without phenotypic selection. Meth. Enzymol. 154, 367–382 (1987).

    Article  CAS  Google Scholar 

  19. Leeuw, T. et al. Pheromone response in yeast: association of Bem1p with proteins of the MAP kinase cascade and actin. Science 270, 1210–1213 (1995).

    Article  ADS  CAS  Google Scholar 

  20. Manser, E., Leung, T., Salihuddin, H., Zhao, Z. S. & Lim, L. Abrain serine/threonine protein kinase activated by Cdc42 and Rac1. Nature 367, 40–46 (1994).

    Article  ADS  CAS  Google Scholar 

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Acknowledgements

We thank A. Nantel for helpful discussion, D. Harcus, D. Dignard and L. Johnson for excellent technical assistance, R. Tsien for supplying the GFP plasmid and S. Sprang for providing the Gβ1 structure coordinates. The NRC publication number for this work is 39976.

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Authors and Affiliations

  1. Eukaryotic Genetics Group, National Research Council of Canada, 6100 Royalmount Avenue, Montreal, Quebec, H4P 2R2, Canada

    Thomas Leeuw, Cunle Wu, Malcolm Whiteway, David Y. Thomas & Ekkehard Leberer

  2. Macromolecular Structure Group, Biotechnology Research Institute, National Research Council of Canada, 6100 Royalmount Avenue, Montreal, Quebec, H4P 2R2, Canada

    Joseph D. Schrag

  3. Department of Biology, McGill University, Montreal, Quebec, H3A 2B2, Canada

    Malcolm Whiteway & David Y. Thomas

  4. Department of Anatomy and Cell Biology, McGill University, Montreal, Quebec, H3A 2B2, Canada

    David Y. Thomas

  5. Department of Medicine, McGill University, Montreal, Quebec, H3A 2B2, Canada

    Ekkehard Leberer

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  1. Thomas Leeuw
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Correspondence to Ekkehard Leberer.

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Leeuw, T., Wu, C., Schrag, J. et al. Interaction of a G-protein β-subunit with a conserved sequence in Ste20/PAK family protein kinases. Nature 391, 191–195 (1998). https://doi.org/10.1038/34448

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