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The MAPK kinase Pek1 acts as a phosphorylation-dependent molecular switch

Abstract

The mitogen-activated protein kinase (MAPK) pathway is a highlyconserved eukaryotic signalling cascade that converts extracellular signals into various outputs, such as cell growth and differentiation1,2,3. MAPK is phosphorylated and activated by a specific MAPK kinase (MAPKK)4: MAPKK is therefore considered to be an activating regulator of MAPK. Pmk1 is a MAPK that regulates cell integrity5 and which, with calcineurin phosphatase, antagonizes chloride homeostasis6 in fission yeast. We have now identified Pek1, a MAPKK for Pmk1 MAPK. We show here that Pek1, in its unphosphorylated form, acts as a potent negative regulator of Pmk1 MAPK signalling. Mkh17, an upstream MAPKK kinase (MAPKKK), converts Pek1 from being an inhibitor to an activator. Our results indicate that Pek1 has a dual stimulatory and inhibitory function which depends on its phosphorylation state. This switch-like mechanism could contribute to the all-or-none physiological response mediated by the MAPK signalling pathway.

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Figure 1: Isolation of Pek1.
Figure 2: Pek1 is a MAPKK for Pmk1.
Figure 3: Pek1 has both an inhibitory and stimulatory effect on Pmk1 MAPK signalling.
Figure 4: The dual function of Pek1 depends on its phosphorylation state.
Figure 5: Mkh1 converts Pek1 from an inhibitor to an activator.
Figure 6: The inhibitory effect of Pek1 on Pmk1 MAPK signalling.

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References

  1. Herskowitz, I. MAP kinase pathways in yeast: for mating and more. Cell 80, 187–197 (1995).

    Article  CAS  Google Scholar 

  2. Nishida, E. & Gotoh, Y. The MAP kinase cascade is essential for diverse signal transduction pathways. Trends Biochem. Sci. 18, 128–131 (1993).

    Article  CAS  Google Scholar 

  3. Levin, D. E. & Errede, B. The proliferation of MAP kinase signaling pathways in yeast. Curr. Biol. 7, 197–202 (1995).

    Article  CAS  Google Scholar 

  4. Marshall, C. J. MAP kinase kinase kinase, MAP kinase kinase and MAP kinase. Curr. Opin. Genet. Dev. 4, 82–89 (1994).

    Article  CAS  Google Scholar 

  5. Toda, T.et al. The fission yeast pmk1+ gene encodes a novel mitogen-activated protein kinase homolog which regulates cell integrity and functions coordinately with the protein kinase C pathway. Mol. Cell. Biol. 16, 6752–6764 (1996).

    Article  CAS  Google Scholar 

  6. Sugiura, R.et al. pmp1+, a suppressor of calcineurin deficiency, encodes a novel MAP kinase phosphatase in fission yeast. EMBO J. 17, 140–148 (1998).

    Article  CAS  Google Scholar 

  7. Sengar, A. S., Markley, N. A., Marini, N. J. & Young, D. Mkh1, a MEK kinase required for cell wall integrity and proper response to osmotic and temperature stress in Schizosaccharomyces pombe. Mol. Cell. Biol. 17, 3508–3519 (1997).

    Article  CAS  Google Scholar 

  8. Yoshida, T., Toda, T. & Yanagida, M. Acalcineurin-like gene ppb1+ in fission yeast: mutant defects in cytokinesis, cell polarity, mating and spindle pole body positioning. J. Cell Sci. 107, 1725–1735 (1994).

    CAS  PubMed  Google Scholar 

  9. Crews, C. M., Alessandrini, A. & Erikson, R. L. The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product. Science 258, 478–480 (1992).

    Article  ADS  CAS  Google Scholar 

  10. Irie, K.et al. MKK1 and MKK2, which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs, function in the pathway mediated by protein kinase C. Mol. Cell. Biol. 13, 3076–3083 (1993).

    Article  CAS  Google Scholar 

  11. Toda, T., Shimanuki, M. & Yanagida, M. Fission yeast genes that confer resistance to staurosporine encode an AP-1-like transcription factor and a protein kinase related to the mammalian ERK1/MAP2 and budding yeast FUS3 and KSS1 kinases. Genes Dev. 5, 60–73 (1991).

    Article  CAS  Google Scholar 

  12. Millar, J. B., Buck, V. & Wilkinson, M. G. Pyp1 and Pyp2 PTPases dephosphorylate an osmosensing MAP kinase controlling cell size at division in fission yeast. Genes Dev. 9, 2117–2130 (1995).

    Article  CAS  Google Scholar 

  13. Shiozaki, K. & Russell, P. Cell-cycle control linked to extracellular environment by MAP kinase pathway in fission yeast. Nature 378, 739–743 (1995).

    Article  ADS  CAS  Google Scholar 

  14. Kato, T.et al. Stress signal, mediated by a Hog1-like MAP kinase, controls sexual development in fission yeast. FEBS Lett. 378, 207–212 (1996).

    Article  CAS  Google Scholar 

  15. Zaitsevskaya, C. T. & Cooper, J. A. Spm1, a stress-activated MAP kinase that regulates morphogenesis in S. pompe. EMBO J. 16, 1318–1331 (1997).

    Article  Google Scholar 

  16. McLeod, M., Stein, M. & Beach, D. The product of the mei3+ gene, expressed under control of the mating-type locus, induces meiosis and sporulation in fission yeast. EMBO J. 6, 729–736 (1987).

    Article  CAS  Google Scholar 

  17. Cook, J. G., Bardwell, L. & Thorner, J. Inhibitory and activating functions for MAPK Kss1 in the S. cerevisiae filamentous-growth signalling pathway. Nature 390, 85–88 (1997).

    Article  ADS  CAS  Google Scholar 

  18. Madhani, H. D., Styles, C. A. & Fink, G. R. MAP kinases with distinct inhibitory functions impart signaling specificity during yeast differentiation. Cell 91, 673–684 (1997).

    Article  CAS  Google Scholar 

  19. Murray, A. W. MAP kinases in meiosis. Cell 92, 157–159 (1998).

    Article  CAS  Google Scholar 

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

    Article  ADS  CAS  Google Scholar 

  21. Rothstein, R. J. One-step gene disruption in yeast. Methods Enzymol. 101, 202–211 (1983).

    Article  CAS  Google Scholar 

  22. Keeney, J. B. & Boeke, J. D. Efficient targeted integration at leu1-32 and ura4-294 in Schizosaccharomyces pombe. Genetics 136, 849–856 (1994).

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Neiman, A. M. & Herskowitz, I. Reconstitution of a yeast protein kinase cascade in vitro: Activation of the yeast MEK homologue STE7 by STE11. Proc. Natl Acad. Sci. USA 91, 3398–3402 (1994).

    Article  ADS  CAS  Google Scholar 

  24. Zarzov, P., Mazzoni, C. & Mann, C. The SLT2(MPK1) MAP kinase is activated during periods of polarized cell growth in yeast. EMBO J. 15, 83–91 (1996).

    Article  CAS  Google Scholar 

  25. Beach, D., Piper, M. & Nurse, P. Construction of a Schizosaccharomyces pombe gene bank in a yeast bacterial shuttle vector and its use to isolate genes by complementation. Mol. Gen. Genet. 187, 326–329 (1982).

    Article  CAS  Google Scholar 

  26. Levin, D. E. & Bishop, M. Aputative protein kinase gene (kin1+) is important for growth polarity in Schizosaccharomyces pombe. Proc. Natl Acad. Sci. USA 87, 8272–8276 (1990).

    Article  ADS  CAS  Google Scholar 

  27. Barbet, N., Muriel, W. J. & Carr, A. M. Versatile shuttle vectors and genomic libraries for use with Schizosaccharomyces pombe. Gene 114, 59–66 (1992).

    Article  CAS  Google Scholar 

  28. Moreno, S., Klar, A. & Nurse, P. Molecular genetic analysis of fission yeast Schizosaccharomyces pombe. Methods Enzymol. 194, 795–823 (1991).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank S. O. Sio, Y. Minami and K. Tanaka for comments on the manuscript, M.Yanagida and H. Mukai for helpful suggestions, and Y. Kawabe for technical assistance. This work was supported in part by research grants from the Ministry of Education, Science and Culture of Japan.

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Sugiura, R., Toda, T., Dhut, S. et al. The MAPK kinase Pek1 acts as a phosphorylation-dependent molecular switch. Nature 399, 479–483 (1999). https://doi.org/10.1038/20951

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