Mae mediates MAP kinase phosphorylation of Ets transcription factors in Drosophila

Abstract

The evolutionarily conserved Ras/mitogen-activated protein kinase (MAPK) cascade is an integral part of the processes of cell division, differentiation, movement and death. Signals received at the cell surface are relayed into the nucleus, where MAPK phosphorylates and thereby modulates the activities of a subset of transcription factors1,2. Here we report the cloning and characterization of a new component of this signal transduction pathway called Mae (for modulator of the activity of Ets). Mae is a signalling intermediate that directly links the MAPK signalling pathway to its downstream transcription factor targets. Phosphorylation by MAPK of the critical serine residue (Ser 127) of the Drosophila transcription factor Yan depends on Mae, and is mediated by the binding of Yan to Mae through their Pointed domains. This phosphorylation is both necessary and sufficient to abrogate transcriptional repression by Yan. Mae also regulates the activity of the transcriptional activator Pointed-P2 by a similar mechanism. Mae is essential for the normal development and viability of Drosophila, and is required in vivo for normal signalling of the epidermal growth factor receptor. Our study indicates that MAPK signalling specificity may depend on proteins that couple specific substrates to the kinase.

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Figure 1: mae sequence and gene organization.
Figure 2: The physical association of Yan and Mae is mediated by their Pnt domains, and this inhibits Yan from binding to DNA.
Figure 3: Mae allows Erk kinase to phosphorylate critical serine or threonine residues of Yan and Pnt-P2, and thus regulates Ets transcriptional activity.
Figure 4: Evidence that mae regulates EGFR signalling in the Drosophila embryonic neurectoderm.

Accession codes

Accessions

GenBank/EMBL/DDBJ

Data deposits

The sequence has been deposited in GenBank under accession number AF358670.

References

  1. 1

    Hill, C. S. & Treisman, R. Transcriptional regulation by extracellular signals: mechanisms and specificity. Cell 80, 199– 211 (1995).

  2. 2

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

  3. 3

    Treisman, R. Regulation of transcription by MAP kinase cascades. Curr. Opin. Cell Biol. 8, 205– 215 (1996).

  4. 4

    Wasylyk, B., Hagman, J. & Gutierrez-Hartmann, A. Ets transcription factors: nuclear effectors of the Ras-MAP-kinase signaling pathway. Trends Biochem. Sci. 23, 213– 216 (1998).

  5. 5

    Lai, Z. C. & Rubin, G. M. Negative control of photoreceptor development in Drosophila by the product of the yan gene, an ETS domain protein. Cell 70, 609– 620 (1992).

  6. 6

    Rebay, I. & Rubin, G. M. Yan functions as a general inhibitor of differentiation and is negatively regulated by activation of the Ras1/MAPK pathway. Cell 80, 857– 866 (1995).

  7. 7

    Rogge, R. et al. The role of yan in mediating the choice between cell division and differentiation. Development 121, 3947– 3958 (1995).

  8. 8

    Rebay, I. et al. A genetic screen for novel components of the Ras/mitogen-activated protein kinase signalling pathway that interact with the yan gene of Drosophila identifies split ends, a new RNA recognition motif-containing protein. Genetics 154, 695– 712 (2000).

  9. 9

    O'Neill, E. M., Rebay, I., Tjian, R. & Rubin, G. M. The activities of two ETS-related transcription factors required for Drosophila eye development are modulated by the Ras/MAPK pathway. Cell 78, 137– 147 (1994).

  10. 10

    Jousset, C. et al. A domain of TEL conserved in a subset of ETS proteins defines a specific oligomerization interface essential to the mitogenic properties of the TEL- PDGFRb oncoprotein. EMBO J. 16, 69– 82 (1997).

  11. 11

    Ponting, C. P. SAM: a novel motif in yeast sterile and Drosophila polyhomeotic proteins. Protein Sci. 4, 1928– 1930 (1995).

  12. 12

    Nye, J. A. Interaction of murine ETS-1 with GGA-binding sites establishes the ETS domain as a new DNA-binding motif. Genes Dev. 6, 975– 990 (1992).

  13. 13

    Soudant, S. et al. A residue in the ETS domain mutated in the v-ets oncogene is essential for the DNA-binding and transactivating properties of the ETS-1 and ETS-2 proteins. Nucleic Acids Res. 22, 3871– 3879 (1994).

  14. 14

    Klambt, C. The Drosophila gene pointed encodes two ETS-like proteins which are involved in the development of midline glial cells. Development 117, 163– 176 (1993).

  15. 15

    Brunner, D. et al. The ETS domain protein Pointed-P2 is a target of MAP kinase in the sevenless signal transduction pathway. Nature 370, 386– 389 (1994).

  16. 16

    Golembo, M., Raz, E. & Shilo, B.-Z. The Drosophila embryonic midline is the site of Spitz processing, and induces activation of the EGF receptor in the ventral ectoderm. Development 122, 3363– 3370 (1996).

  17. 17

    Yagi, Y., Suzuki, T. & Hayashi, S. Interaction between Drosophila EGF receptor and vnd determines three dorsoventral domains of the neurectoderm. Development 125, 3625– 3633 (1998).

  18. 18

    Gabay, L. et al. EGF receptor signalling induces pointed P1 transcription and inactivates Yan protein in the Drosophila embryonic ventral ectoderm. Development 122, 3355– 3362 (1996).

  19. 19

    Golembo, M., Yarnitzky, T., Volk, T. & Shilo, B.-Z. Vein expression is induced by the EGF receptor pathway to provide a positive feedback loop in patterning the Drosophila embryonic ventral ectoderm. Genes Dev. 13, 158– 162 (1999).

  20. 20

    Gabay, L., Seger, R. & Shilo, B.-Z. In situ activation pattern of Drosophila EGF receptor pathway during development. Science 77, 1103– 1106 (1997).

  21. 21

    Dumstrei, K. et al. EGFR signalling is required for the differentiation and maintenance of neural progenitors along the dorsal midline of the Drosophila embryonic head. Development 125, 3417– 3426 (1998).

  22. 22

    Roch, F. et al. Screening of larval/pupal P-element induced lethals on the second chromosome in Drosophila melanogaster: clonal analysis and morphology of imaginal discs. Mol. Gen. Genet. 257, 103– 112 (1998).

  23. 23

    Spradling, A. C. et al. The Berkeley Drosophila genome gene disruption project. Single P-element insertions mutating 25% of vital Drosophila genes. Genetics 153, 135– 177 (1999).

  24. 24

    Jurgens, G., Wieschaus, E., Nusslein-Volhard, C. & Kluding, H. Mutations affecting the pattern of the larval cuticle in Drosophila melanogaster II. Zygotic loci on the third chromosome. Roux's Arch. Dev. Biol. 193, 283– 295 (1984).

  25. 25

    Mayer, U. & Nusselein-Volhard, C. A group of genes required for pattern formation in the ventral ectoderm of the Drosophila embryo. Genes Dev. 2, 1496– 1503 (1988).

  26. 26

    Wasserman, J. D., Urban, S. & Freeman, M. A family of rhomboid-like genes: Drosophila rhomboid-1 and roughoid/rhomboid-3 cooperate to activate EGF receptor signalling. Genes Dev. 14, 1651– 1663 (2000).

  27. 27

    Golembo, M., Schweitzer, R., Freeman, M. & Shilo, B. -Z. Argos transcription is induced by the Drosophila EGF receptor pathway to form an inhibitory feedback loop. Development 122, 223– 230 (1996).

  28. 28

    Scholz, H., Sadlowski, E., Klaes, A. & Klambt, C. Control of midline glia development in the embryonic Drosophila CNS. Mech. Dev. 64, 137– 151 (1997).

  29. 29

    Towbin, H., Staehelin, T. & Gordon, J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl Acad. Sci. USA 76, 4350– 4354 (1979).

  30. 30

    Wieschaus, E. & Nusslein-Volhard, C. in Drosophila, a Practical Approach (ed. Roberts, D. B) 199– 226 (IRL, Oxford, 1986).

  31. 31

    Tautz, D. & Pfeifle, C. A non-radioactive in situ hybridization method for the localization of specific RNAs in Drosophila embryos reveals translational control of the segmentation gene hunchback. Chromosoma 98, 81– 85 (1989).

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Acknowledgements

We thank S. Bullock, R. Winston, H. McNeill, P. Verrijzer, C. Hill, A. Maata, P. Mason, J. White, J. Brossens, M. Freeman, G. Rubin, T. Laverty and P. Soccorso for advice, discussions and gifts of materials. This work was supported by the Imperial Cancer Research Fund and BBSRC.

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Correspondence to David A. Baker.

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Baker, D., Mille-Baker, B., Wainwright, S. et al. Mae mediates MAP kinase phosphorylation of Ets transcription factors in Drosophila. Nature 411, 330–334 (2001). https://doi.org/10.1038/35077122

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