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. 1996 Sep 2;15(17):4556–4565.

Regulation of embryonic cell division by a Xenopus gastrula-specific protein kinase.

A M Snape 1, J C Smith 1
PMCID: PMC452185  PMID: 8887547

Abstract

We describe a novel protein kinase, Pk9.7, and its role in cell division in the Xenopus embryo. Pk9.7 is transcribed only during blastula and gastrula stages. Expression of Pk9.7 in Xenopus oocytes induces meiotic maturation, while overexpression in embryos blocks blastomere cleavage in a MAP kinase-independent fashion. In both Pk9.7-injected oocytes and mitotic cells of cleavage-blocked embryos, chromosomes appear detached from abnormal spindles, and in oocytes additional microtubule structures are formed, suggesting that one function of Pk9.7 is to regulate formation of, and chromosome attachment to, the spindle. Consistent with this, Pk9.7 co-immunoprecipitates tubulin and phosphorylates it in vitro. Pk9.7 expression coincides with the switch from maternal to zygotic control of the cell cycle, and with the switch from microtubule independence to microtubule dependence. Our results suggest that Pk9.7 plays a role in these processes.

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Selected References

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  1. Almouzni G., Wolffe A. P. Constraints on transcriptional activator function contribute to transcriptional quiescence during early Xenopus embryogenesis. EMBO J. 1995 Apr 18;14(8):1752–1765. doi: 10.1002/j.1460-2075.1995.tb07164.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Campbell M. S., Gorbsky G. J. Microinjection of mitotic cells with the 3F3/2 anti-phosphoepitope antibody delays the onset of anaphase. J Cell Biol. 1995 Jun;129(5):1195–1204. doi: 10.1083/jcb.129.5.1195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cho K. W., Blumberg B., Steinbeisser H., De Robertis E. M. Molecular nature of Spemann's organizer: the role of the Xenopus homeobox gene goosecoid. Cell. 1991 Dec 20;67(6):1111–1120. doi: 10.1016/0092-8674(91)90288-a. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clute P., Masui Y. Regulation of the appearance of division asynchrony and microtubule-dependent chromosome cycles in Xenopus laevis embryos. Dev Biol. 1995 Oct;171(2):273–285. doi: 10.1006/dbio.1995.1280. [DOI] [PubMed] [Google Scholar]
  5. Cunliffe V., Smith J. C. Specification of mesodermal pattern in Xenopus laevis by interactions between Brachyury, noggin and Xwnt-8. EMBO J. 1994 Jan 15;13(2):349–359. doi: 10.1002/j.1460-2075.1994.tb06268.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Daar I., Nebreda A. R., Yew N., Sass P., Paules R., Santos E., Wigler M., Vande Woude G. F. The ras oncoprotein and M-phase activity. Science. 1991 Jul 5;253(5015):74–76. doi: 10.1126/science.1829549. [DOI] [PubMed] [Google Scholar]
  7. Daar I., Yew N., Vande Woude G. F. Inhibition of mos-induced oocyte maturation by protein kinase A. J Cell Biol. 1993 Mar;120(5):1197–1202. doi: 10.1083/jcb.120.5.1197. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dumont J. N. Oogenesis in Xenopus laevis (Daudin). I. Stages of oocyte development in laboratory maintained animals. J Morphol. 1972 Feb;136(2):153–179. doi: 10.1002/jmor.1051360203. [DOI] [PubMed] [Google Scholar]
  9. Edgar B. A., Lehman D. A., O'Farrell P. H. Transcriptional regulation of string (cdc25): a link between developmental programming and the cell cycle. Development. 1994 Nov;120(11):3131–3143. doi: 10.1242/dev.120.11.3131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610-3616.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gotoh Y., Masuyama N., Suzuki A., Ueno N., Nishida E. Involvement of the MAP kinase cascade in Xenopus mesoderm induction. EMBO J. 1995 Jun 1;14(11):2491–2498. doi: 10.1002/j.1460-2075.1995.tb07246.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gotoh Y., Nishida E. The MAP kinase cascade: its role in Xenopus oocytes, eggs and embryos. Prog Cell Cycle Res. 1995;1:287–297. doi: 10.1007/978-1-4615-1809-9_23. [DOI] [PubMed] [Google Scholar]
  13. Gotoh Y., Nishida E., Yamashita T., Hoshi M., Kawakami M., Sakai H. Microtubule-associated-protein (MAP) kinase activated by nerve growth factor and epidermal growth factor in PC12 cells. Identity with the mitogen-activated MAP kinase of fibroblastic cells. Eur J Biochem. 1990 Nov 13;193(3):661–669. doi: 10.1111/j.1432-1033.1990.tb19384.x. [DOI] [PubMed] [Google Scholar]
  14. Graves L. M., Northrop J. L., Potts B. C., Krebs E. G., Kimelman D. Fibroblast growth factor, but not activin, is a potent activator of mitogen-activated protein kinase in Xenopus explants. Proc Natl Acad Sci U S A. 1994 Mar 1;91(5):1662–1666. doi: 10.1073/pnas.91.5.1662. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Green J. B., New H. V., Smith J. C. Responses of embryonic Xenopus cells to activin and FGF are separated by multiple dose thresholds and correspond to distinct axes of the mesoderm. Cell. 1992 Nov 27;71(5):731–739. doi: 10.1016/0092-8674(92)90550-v. [DOI] [PubMed] [Google Scholar]
  16. Gurdon J. B., Brennan S., Fairman S., Mohun T. J. Transcription of muscle-specific actin genes in early Xenopus development: nuclear transplantation and cell dissociation. Cell. 1984 Oct;38(3):691–700. doi: 10.1016/0092-8674(84)90264-2. [DOI] [PubMed] [Google Scholar]
  17. Haccard O., Sarcevic B., Lewellyn A., Hartley R., Roy L., Izumi T., Erikson E., Maller J. L. Induction of metaphase arrest in cleaving Xenopus embryos by MAP kinase. Science. 1993 Nov 19;262(5137):1262–1265. doi: 10.1126/science.8235656. [DOI] [PubMed] [Google Scholar]
  18. Hanks S. K., Quinn A. M., Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988 Jul 1;241(4861):42–52. doi: 10.1126/science.3291115. [DOI] [PubMed] [Google Scholar]
  19. Houliston E., Elinson R. P. Patterns of microtubule polymerization relating to cortical rotation in Xenopus laevis eggs. Development. 1991 May;112(1):107–117. doi: 10.1242/dev.112.1.107. [DOI] [PubMed] [Google Scholar]
  20. Howe J. A., Howell M., Hunt T., Newport J. W. Identification of a developmental timer regulating the stability of embryonic cyclin A and a new somatic A-type cyclin at gastrulation. Genes Dev. 1995 May 15;9(10):1164–1176. doi: 10.1101/gad.9.10.1164. [DOI] [PubMed] [Google Scholar]
  21. Isaacs H. V., Tannahill D., Slack J. M. Expression of a novel FGF in the Xenopus embryo. A new candidate inducing factor for mesoderm formation and anteroposterior specification. Development. 1992 Mar;114(3):711–720. doi: 10.1242/dev.114.3.711. [DOI] [PubMed] [Google Scholar]
  22. King R. W., Jackson P. K., Kirschner M. W. Mitosis in transition. Cell. 1994 Nov 18;79(4):563–571. doi: 10.1016/0092-8674(94)90542-8. [DOI] [PubMed] [Google Scholar]
  23. Krieg P. A., Melton D. A. Functional messenger RNAs are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res. 1984 Sep 25;12(18):7057–7070. doi: 10.1093/nar/12.18.7057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. LaBonne C., Burke B., Whitman M. Role of MAP kinase in mesoderm induction and axial patterning during Xenopus development. Development. 1995 May;121(5):1475–1486. doi: 10.1242/dev.121.5.1475. [DOI] [PubMed] [Google Scholar]
  25. MacNicol A. M., Muslin A. J., Howard E. L., Kikuchi A., MacNicol M. C., Williams L. T. Regulation of Raf-1-dependent signaling during early Xenopus development. Mol Cell Biol. 1995 Dec;15(12):6686–6693. doi: 10.1128/mcb.15.12.6686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. MacNicol A. M., Muslin A. J., Williams L. T. Raf-1 kinase is essential for early Xenopus development and mediates the induction of mesoderm by FGF. Cell. 1993 May 7;73(3):571–583. doi: 10.1016/0092-8674(93)90143-e. [DOI] [PubMed] [Google Scholar]
  27. Maller J. L., Krebs E. G. Progesterone-stimulated meiotic cell division in Xenopus oocytes. Induction by regulatory subunit and inhibition by catalytic subunit of adenosine 3':5'-monophosphate-dependent protein kinase. J Biol Chem. 1977 Mar 10;252(5):1712–1718. [PubMed] [Google Scholar]
  28. Meyerson M., Enders G. H., Wu C. L., Su L. K., Gorka C., Nelson C., Harlow E., Tsai L. H. A family of human cdc2-related protein kinases. EMBO J. 1992 Aug;11(8):2909–2917. doi: 10.1002/j.1460-2075.1992.tb05360.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Minshull J., Sun H., Tonks N. K., Murray A. W. A MAP kinase-dependent spindle assembly checkpoint in Xenopus egg extracts. Cell. 1994 Nov 4;79(3):475–486. doi: 10.1016/0092-8674(94)90256-9. [DOI] [PubMed] [Google Scholar]
  30. Mohun T., Garrett N., Stutz F., Sophr G. A third striated muscle actin gene is expressed during early development in the amphibian Xenopus laevis. J Mol Biol. 1988 Jul 5;202(1):67–76. doi: 10.1016/0022-2836(88)90519-0. [DOI] [PubMed] [Google Scholar]
  31. Muslin A. J., MacNicol A. M., Williams L. T. Raf-1 protein kinase is important for progesterone-induced Xenopus oocyte maturation and acts downstream of mos. Mol Cell Biol. 1993 Jul;13(7):4197–4202. doi: 10.1128/mcb.13.7.4197-4202.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Nebreda A. R., Hunt T. The c-mos proto-oncogene protein kinase turns on and maintains the activity of MAP kinase, but not MPF, in cell-free extracts of Xenopus oocytes and eggs. EMBO J. 1993 May;12(5):1979–1986. doi: 10.1002/j.1460-2075.1993.tb05847.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Nebreda A. R., Porras A., Santos E. p21ras-induced meiotic maturation of Xenopus oocytes in the absence of protein synthesis: MPF activation is preceded by activation of MAP and S6 kinases. Oncogene. 1993 Feb;8(2):467–477. [PubMed] [Google Scholar]
  34. Newport J., Kirschner M. A major developmental transition in early Xenopus embryos: I. characterization and timing of cellular changes at the midblastula stage. Cell. 1982 Oct;30(3):675–686. doi: 10.1016/0092-8674(82)90272-0. [DOI] [PubMed] [Google Scholar]
  35. Nicklas R. B., Krawitz L. E., Ward S. C. Odd chromosome movement and inaccurate chromosome distribution in mitosis and meiosis after treatment with protein kinase inhibitors. J Cell Sci. 1993 Apr;104(Pt 4):961–973. doi: 10.1242/jcs.104.4.961. [DOI] [PubMed] [Google Scholar]
  36. Posada J., Yew N., Ahn N. G., Vande Woude G. F., Cooper J. A. Mos stimulates MAP kinase in Xenopus oocytes and activates a MAP kinase kinase in vitro. Mol Cell Biol. 1993 Apr;13(4):2546–2553. doi: 10.1128/mcb.13.4.2546-2553.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Rupp R. A., Snider L., Weintraub H. Xenopus embryos regulate the nuclear localization of XMyoD. Genes Dev. 1994 Jun 1;8(11):1311–1323. doi: 10.1101/gad.8.11.1311. [DOI] [PubMed] [Google Scholar]
  38. Sagata N., Oskarsson M., Copeland T., Brumbaugh J., Vande Woude G. F. Function of c-mos proto-oncogene product in meiotic maturation in Xenopus oocytes. Nature. 1988 Oct 6;335(6190):519–525. doi: 10.1038/335519a0. [DOI] [PubMed] [Google Scholar]
  39. Sagata N., Watanabe N., Vande Woude G. F., Ikawa Y. The c-mos proto-oncogene product is a cytostatic factor responsible for meiotic arrest in vertebrate eggs. Nature. 1989 Nov 30;342(6249):512–518. doi: 10.1038/342512a0. [DOI] [PubMed] [Google Scholar]
  40. Sargent M. G., Bennett M. F. Identification in Xenopus of a structural homologue of the Drosophila gene snail. Development. 1990 Aug;109(4):967–973. doi: 10.1242/dev.109.4.967. [DOI] [PubMed] [Google Scholar]
  41. Scavo L., Shuldiner A. R., Serrano J., Dashner R., Roth J., de Pablo F. Genes encoding receptors for insulin and insulin-like growth factor I are expressed in Xenopus oocytes and embryos. Proc Natl Acad Sci U S A. 1991 Jul 15;88(14):6214–6218. doi: 10.1073/pnas.88.14.6214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Smith J. C., Price B. M., Green J. B., Weigel D., Herrmann B. G. Expression of a Xenopus homolog of Brachyury (T) is an immediate-early response to mesoderm induction. Cell. 1991 Oct 4;67(1):79–87. doi: 10.1016/0092-8674(91)90573-h. [DOI] [PubMed] [Google Scholar]
  43. Steele R. E., Irwin M. Y., Knudsen C. L., Collett J. W., Fero J. B. The yes proto-oncogene is present in amphibians and contributes to the maternal RNA pool in the oocyte. Oncogene Res. 1989;4(3):223–233. [PubMed] [Google Scholar]
  44. Thomas B. J., Gunning D. A., Cho J., Zipursky L. Cell cycle progression in the developing Drosophila eye: roughex encodes a novel protein required for the establishment of G1. Cell. 1994 Jul 1;77(7):1003–1014. doi: 10.1016/0092-8674(94)90440-5. [DOI] [PubMed] [Google Scholar]
  45. Toda T., Cameron S., Sass P., Zoller M., Wigler M. Three different genes in S. cerevisiae encode the catalytic subunits of the cAMP-dependent protein kinase. Cell. 1987 Jul 17;50(2):277–287. doi: 10.1016/0092-8674(87)90223-6. [DOI] [PubMed] [Google Scholar]
  46. Umbhauer M., Marshall C. J., Mason C. S., Old R. W., Smith J. C. Mesoderm induction in Xenopus caused by activation of MAP kinase. Nature. 1995 Jul 6;376(6535):58–62. doi: 10.1038/376058a0. [DOI] [PubMed] [Google Scholar]
  47. Winning R. S., Sargent T. D. Pagliaccio, a member of the Eph family of receptor tyrosine kinase genes, has localized expression in a subset of neural crest and neural tissues in Xenopus laevis embryos. Mech Dev. 1994 Jun;46(3):219–229. doi: 10.1016/0925-4773(94)90072-8. [DOI] [PubMed] [Google Scholar]
  48. Yasuda G. K., Schubiger G. Temporal regulation in the early embryo: is MBT too good to be true? Trends Genet. 1992 Apr;8(4):124–127. doi: 10.1016/0168-9525(92)90369-F. [DOI] [PubMed] [Google Scholar]
  49. Yew N., Mellini M. L., Vande Woude G. F. Meiotic initiation by the mos protein in Xenopus. Nature. 1992 Feb 13;355(6361):649–652. doi: 10.1038/355649a0. [DOI] [PubMed] [Google Scholar]
  50. Yi T. L., Bolen J. B., Ihle J. N. Hematopoietic cells express two forms of lyn kinase differing by 21 amino acids in the amino terminus. Mol Cell Biol. 1991 May;11(5):2391–2398. doi: 10.1128/mcb.11.5.2391-2398.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Zhou R. P., Oskarsson M., Paules R. S., Schulz N., Cleveland D., Vande Woude G. F. Ability of the c-mos product to associate with and phosphorylate tubulin. Science. 1991 Feb 8;251(4994):671–675. doi: 10.1126/science.1825142. [DOI] [PubMed] [Google Scholar]

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