Tyrosine Phosphorylation of MPF and Membrane Proteins During Meiotic Maturation of Starfish Oocytes

  • G. Peaucellier
  • A. C. Andersen
  • W. H. Kinsey
  • M. Dorée
Conference paper
Part of the NATO ASI Series book series (volume 45)


It has been reported that the starfish homolog of the fission yeast cdc2 protein is a component of maturation promoting factor (MPF) that controls entry of eukaryotic cells into M-phase. The p34 cdc2 protein is phosphorylated during interphase and dephosphorylated during M-phase. We show that starfish p34 cdc2 is phosphorylated in vivo on tyrosine, threonine and serine in G2 (prophase I) and Gl (after completion of meiotic divisions). Rephosphorylation of p34 cdc2 is not prevented or modified by inhibition of protein synthesis during M-phase.

Tyrosine phosphorylation was found to occur in proteins from oocytes cortices, by immunoprecipitations with an antibody specific for phosphotyrosine. In 32P-preloaded oocytes, labeled phosphotyrosine containing proteins were noticeable only after hormonal induction of meiotic divisions. Labeling increased in five major phosphoproteins of 195, 155, 100, 45 and 35 kDa until first polar body emission, then decreased upon completion of meiosis. Endogenous in vitro phosphorylation of cortices showed a high tyrosine kinase activity towards a 68-kDa protein but no difference between cortices from oocytes treated or not with the hormone.

These findings suggest that tyrosine phosphorylation contributes both to the transduction of the hormonal signal for meiosis resumption and to several steps of the ensuing divisions.


Tyrosine Phosphorylation Fission Yeast Meiotic Division Cortical Granule Meiotic Maturation 
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  1. Arion D, Meijer L, Brizuela L and Beach D (1988) cdc2 is a component of the M phase-specific histone H1 kinase: evidence for identity with MPF. Cell 55:371–378PubMedCrossRefGoogle Scholar
  2. Bourassa C, Chapdelaine A, Roberts K D and Chevalier S (1988) Enhancement of the detection of alkali-resistant phosphoproteins in Polyacrylamide gels. Anal. Biochem. 169:356–362PubMedCrossRefGoogle Scholar
  3. Brizuela L, Draetta G and Beach D (1987) pl3sucl acts in the fission yeast cell division cycle as a component of the p34cdc2 protein kinase. EMBO J. 6:3507–3514PubMedGoogle Scholar
  4. Brizuela L, Draetta G and Beach D (1989) Activation of cdc2 protein as a histone H1 kinase is associated with complex formation with the p62 subunit. Proc. Natl. Acad. Sci. U.S.A. 86:4362–4366PubMedCrossRefGoogle Scholar
  5. Capony J-P, Picard A, Peaucellier G, Labbé J-C and Dorée M (1986) Changes in the activity of the maturation-promoting factor during meiotic maturation and following activation of amphibian and starfish oocytes: their correlation with protein phosphorylation. Dev. Biol. 117:1–12CrossRefGoogle Scholar
  6. Chen WS, Lazar CS, Poenie M, Tsien RY, Gill GN and Rosenfeld MG (1987) Requirement for intrinsic protein tyrosine kinase in the immediate and late actions of the EGF receptor. Nature 328:820–823PubMedCrossRefGoogle Scholar
  7. Chou CK, Dull TJ, Russell DS, Gherzi R, Lebwohl D, Ulrich U and Rosen OM (1987) Human insulin receptors mutated at the ATP-binding site lack protein tyrosine kinase activity and fail to mediate postreceptor effects of insulin. J. Biol. Chem. 262:1842–1847PubMedGoogle Scholar
  8. Cooper JA, Sefton BM and Hunter T (1983) Detection and quantification of phosphotyrosine in proteins. Methods Enzymol. 99:387–401PubMedCrossRefGoogle Scholar
  9. Dasgrupta JD and Garbers DL (1983) Tyrosine protein kinase activity during embryogenesis. J. Biol. Chem. 258:6174–6178Google Scholar
  10. Detering NK, Decker GL, Schmell ED and Lennarz WJ (1977) Isolation and characterization of plasma membrane-associated cortical granules from sea urchin eggs. J. Cell Biol. 75:899–914PubMedCrossRefGoogle Scholar
  11. Dorée M and Guerrier P (1975) Site of action of 1-methyladenine in inducing oocyte maturation in starfishes: Kinetical evidences for receptors localized on the cell surface. Exp. Cell Res. 91:296–300PubMedCrossRefGoogle Scholar
  12. Dorée M, Peaucellier G and Picard A (1983) Activity of the maturation-promoting factor and the extent of protein phosphorylation oscillate simultaneously during meiotic maturation of starfish oocytes. Dev. Biol. 99:489–501PubMedCrossRefGoogle Scholar
  13. Dunphy WG and Newport JW (1988) Unraveling of mitotic control mechanisms. Cell 55:925–928PubMedCrossRefGoogle Scholar
  14. Dunphy WG and Newport JW (1989) Fission yeast pl3 blocks mitotic activation and tyrosine dephosphorylation of the Xenopus cdc2 protein kinase. Cell 58:181–191PubMedCrossRefGoogle Scholar
  15. Draetta G, Piwnica-Worms H, Morrison D, Druker B, Roberts T and Beach D (1988) Human cdc2 protein kinase is a major cell-cycle regulated tyrosine kinase substrate. Nature 336:738–743PubMedCrossRefGoogle Scholar
  16. Gautier J, Matsukawa T, Nurse P and Maller J (1989) Dephosphorylation and activation of “Xenopus” p34cdc2 protein kinase during the cell cycle. Nature 339:626–628PubMedCrossRefGoogle Scholar
  17. Guerrier P and Dorée M (1975) Hormonal control of reinitiation of meiosis in starfish. The requirement of 1-methyladenine during nuclear maturation. Dev. Biol. 47:341–348PubMedCrossRefGoogle Scholar
  18. Heldin CH and Westermark B (1984) Growth factors: mechanism of action and relation to oncogenes. Cell 37:9–20PubMedCrossRefGoogle Scholar
  19. Hunter T and Cooper J.A. (1985) Protein-tyrosine kinases. Annu. Rev. Biochem. 54:897–930PubMedCrossRefGoogle Scholar
  20. Kamel C, Veno PA and Kinsey WH (1986) Quantitation of a src-like tyrosine protein kinase during fertilization of the sea urchin egg. Biochem. Biophys. Res. Commun. 138:349–355PubMedCrossRefGoogle Scholar
  21. Kamps MP and Sefton BM (1988) Identification of multiple novel polypeptide substrates of the v-src, v-yes, v-fps, v-ros, and v-erb-B oncogenic protein kinases utilizing antisera against phosphotyrosine. Oncogene 2:305–315PubMedGoogle Scholar
  22. Kinsey WH (1984) Regulation of tyrosine-specific kinase activity at fertilization. Dev. Biol. 105:137–143PubMedCrossRefGoogle Scholar
  23. Kinsey WH, Decker GL and Lennarz WJ (1980) Isolation and partial characterization of the plasma membrane of the sea urchin egg. J. Cell Biol. 87:248–254PubMedCrossRefGoogle Scholar
  24. Kishimoto T, Kuriyama R, Kondo H and Kanatani H (1982) Generality of the action of various maturation-promoting factors. Exptl. Cell Res. 137: 121–126PubMedCrossRefGoogle Scholar
  25. Labbé J-C, Capony J-P, Caput D, Cavadore J-C, Derancourt J, Kaghad M, Lelias J-M, Picard A and Dorée M (1989) MPF from starfish oocytes at first meiotic metaphase is an heterodimer containing one molecule of cdc2 and one molecule of cyclin B. EMBO J. 8:3053–3058PubMedGoogle Scholar
  26. Labbé J-C, Lee MG, Nurse P, Picard A and Dorée M (1988) Activation at M-phase of a protein kinase encoded by a starfish homologue of the cell cycle control gene cdc2+. Nature 335:251–253PubMedCrossRefGoogle Scholar
  27. Labbé JC, Picard A, Peaucellier G, Lee M, Nurse P and Dorée M (1989) Purification of MPF from starfish: identification as the H1 histone kinase p34cdc2 and a possible mechanism for its periodic activation. Cell 57:253–263PubMedCrossRefGoogle Scholar
  28. Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685PubMedCrossRefGoogle Scholar
  29. Lee M and Nurse P (1988) Cell cycle control genes in fission yeast and mammalian cells. Trends Genet. 4:287PubMedCrossRefGoogle Scholar
  30. Meijer L, Arion D, Goldstein R, Pines J, Brizuela L, Hunt T and Beach D (1989) Cyclin is a component of the sea urchin egg M-phase specific histone H1 kinase. EMBO J. 8:2275–2282PubMedGoogle Scholar
  31. Moria AO, Draetta G, Beach D and Wang JY (1989) Reversible tyrosine phosphorylation of cdc2: dephosphorylation accompanies activation during entry into mitosis. Cell 58:193–203)CrossRefGoogle Scholar
  32. Peaucellier G, Veno PA and Einsey WH (1988) Protein tyrosine phosphorylation in response to fertilization. J. Biol. Chem. 263:3806–13811Google Scholar
  33. Picard A, Peaucellier G, Le Bouffant F, Le Peuch C and Dorée M (1985) Role of protein synthesis and proteases in production and inactivation of maturation-promoting activity during meiotic maturation of starfish oocytes. Dev. Biol. 109:311–320PubMedCrossRefGoogle Scholar
  34. Pondaven P, Meijer L and Pelech SL (1987) Protein phosphorylation in starfish oocyte meiotic divisions and sea urchin egg mitotic divisions. Adv. Protein Phosphatases 4:229–251Google Scholar
  35. Ribot HD, Eisenman EA and Kinsey WH (1984) Fertilization results in increased tyrosine phosphorylation of egg proteins. J. Biol. Chem. 259:5333–5338PubMedGoogle Scholar
  36. Ross AH, Baltimore D and Eisen HN (1981) Phosphotyrosine-containing proteins isolated by affinity chromatography with antibodies to a synthetic hapten. Nature 294:654–656PubMedCrossRefGoogle Scholar
  37. Sakanoue Y, Hashimoto E, Nakamura S-I and Yamamura H (1988) Insulin-stimulated serine kinase in Xenopus oocyte plasma membrane. Biochem. Biophys. Res. Commun. 150:1176–1184PubMedCrossRefGoogle Scholar
  38. Satoh N and Garbers DL (1985) Protein tyrosine kinase activity of eggs of the sea urchin Strongylocentrotus purpuratus: The regulation of its increase after fertilization. Dev. Biol. 111:515–519PubMedCrossRefGoogle Scholar
  39. Spivack JG, Erikson RL and Mailer JL (1984) Microinjection of pp60v-src into Xenopus oocytes increases phosphorylation of ribosomal protein S6 and accelerates the rate of progesterone-induced meiotic maturation. Mol. Cell Biol. 4:1631–1634PubMedGoogle Scholar
  40. Wang JYJ (1988) Antibodies for phosphotyrosine: analytical and preparative tool for tyrosyl-phosphorylated proteins. Anal. Biochem. 172:1–7PubMedCrossRefGoogle Scholar
  41. Yarden Y and Ullrich A (1988) Growth factor receptor tyrosine kinases. Ann. Rev. Biochem. 57:443–478PubMedCrossRefGoogle Scholar
  42. Yoshikuni M, Ishikawa K, Isobe M, Goto T and Nagahama Y (1988b) Characterization of 1-methyladenine binding in starfish oocyte cortices. Proc. Natl. Acad. Sci. U.S.A. 85:1874–1877PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 1990

Authors and Affiliations

  • G. Peaucellier
    • 1
  • A. C. Andersen
    • 1
  • W. H. Kinsey
    • 1
    • 2
  • M. Dorée
    • 1
    • 3
  1. 1.Station BiologiqueRoscoffFrance
  2. 2.Department of Anatomy and Cell BiologyUniversity of Miami School of MedicineMiamiUSA
  3. 3.CNRS and INSERMMontpellier CedexFrance

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