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. 1990 Jun;1(7):543–554. doi: 10.1091/mbc.1.7.543

H-ras(val12) induces cytoplasmic but not nuclear events of the cell cycle in small Xenopus oocytes.

A D Johnson 1, R J Cork 1, M A Williams 1, K R Robinson 1, L D Smith 1
PMCID: PMC361572  PMID: 2098115

Abstract

Microinjection of H-ras(val12) protein into fully grown Xenopus oocytes has been shown to induce meiotic maturation. In the present study, mRNA encoding the mutant ras protein was injected into both fully grown (stage 6) and growing (stage 4) oocytes. The mRNA induced nuclear breakdown in stage 6 oocytes, as expected. However, the mRNA induced neither nuclear breakdown nor maturation promoting factor when injected into stage 4 oocytes. Instead, the response in stage 4 oocytes included an activation pulse of calcium, cortical granule breakdown, elevation of the vitelline envelope, and abortive cleavage furrows, all of which are characteristics of the activation response in mature eggs. In addition, the injected mRNA led to increased rates of endogenous protein synthesis and the migration of subcortical organelles into the oocyte interior. These observations are discussed relative to the suggestion that oncogenic ras protein leads to an increase in both diacylglycerol and inositol trisphosphate, which then regulate the various cytoplasmic events described.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allende C. C., Hinrichs M. V., Santos E., Allende J. E. Oncogenic ras protein induces meiotic maturation of amphibian oocytes in the presence of protein synthesis inhibitors. FEBS Lett. 1988 Jul 18;234(2):426–430. doi: 10.1016/0014-5793(88)80130-3. [DOI] [PubMed] [Google Scholar]
  2. Barbacid M. ras genes. Annu Rev Biochem. 1987;56:779–827. doi: 10.1146/annurev.bi.56.070187.004023. [DOI] [PubMed] [Google Scholar]
  3. Barrett C. B., Schroetke R. M., Van der Hoorn F. A., Nordeen S. K., Maller J. L. Ha-rasVal-12,Thr-59 activates S6 kinase and p34cdc2 kinase in Xenopus oocytes: evidence for c-mosxe-dependent and -independent pathways. Mol Cell Biol. 1990 Jan;10(1):310–315. doi: 10.1128/mcb.10.1.310. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bement W. M., Capco D. G. Protein kinase C acts downstream of calcium at entry into the first mitotic interphase of Xenopus laevis. Cell Regul. 1990 Feb;1(3):315–326. doi: 10.1091/mbc.1.3.315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Berridge M. J., Galione A. Cytosolic calcium oscillators. FASEB J. 1988 Dec;2(15):3074–3082. doi: 10.1096/fasebj.2.15.2847949. [DOI] [PubMed] [Google Scholar]
  6. Birchmeier C., Broek D., Wigler M. ras proteins can induce meiosis in Xenopus oocytes. Cell. 1985 Dec;43(3 Pt 2):615–621. doi: 10.1016/0092-8674(85)90233-8. [DOI] [PubMed] [Google Scholar]
  7. Busa W. B., Ferguson J. E., Joseph S. K., Williamson J. R., Nuccitelli R. Activation of frog (Xenopus laevis) eggs by inositol trisphosphate. I. Characterization of Ca2+ release from intracellular stores. J Cell Biol. 1985 Aug;101(2):677–682. doi: 10.1083/jcb.101.2.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Busa W. B., Nuccitelli R. An elevated free cytosolic Ca2+ wave follows fertilization in eggs of the frog, Xenopus laevis. J Cell Biol. 1985 Apr;100(4):1325–1329. doi: 10.1083/jcb.100.4.1325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Charbonneau M., Grey R. D. The onset of activation responsiveness during maturation coincides with the formation of the cortical endoplasmic reticulum in oocytes of Xenopus laevis. Dev Biol. 1984 Mar;102(1):90–97. doi: 10.1016/0012-1606(84)90177-5. [DOI] [PubMed] [Google Scholar]
  10. Cork R. J., Cicirelli M. F., Robinson K. R. A rise in cytosolic calcium is not necessary for maturation of Xenopus laevis oocytes. Dev Biol. 1987 May;121(1):41–47. doi: 10.1016/0012-1606(87)90136-9. [DOI] [PubMed] [Google Scholar]
  11. Deshpande A. K., Kung H. F. Insulin induction of Xenopus laevis oocyte maturation is inhibited by monoclonal antibody against p21 ras proteins. Mol Cell Biol. 1987 Mar;7(3):1285–1288. doi: 10.1128/mcb.7.3.1285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Fasano O., Taparowsky E., Fiddes J., Wigler M., Goldfarb M. Sequence and structure of the coding region of the human H-ras-1 gene from T24 bladder carcinoma cells. J Mol Appl Genet. 1983;2(2):173–180. [PubMed] [Google Scholar]
  14. Ford C. C., Gurdon J. B. A method for enucleating oocytes of Xenopus laevis. J Embryol Exp Morphol. 1977 Feb;37(1):203–209. [PubMed] [Google Scholar]
  15. Galili G., Kawata E. E., Smith L. D., Larkins B. A. Role of the 3'-poly(A) sequence in translational regulation of mRNAs in Xenopus laevis oocytes. J Biol Chem. 1988 Apr 25;263(12):5764–5770. [PubMed] [Google Scholar]
  16. Gerhart J., Wu M., Kirschner M. Cell cycle dynamics of an M-phase-specific cytoplasmic factor in Xenopus laevis oocytes and eggs. J Cell Biol. 1984 Apr;98(4):1247–1255. doi: 10.1083/jcb.98.4.1247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gibbs J. B., Schaber M. D., Schofield T. L., Scolnick E. M., Sigal I. S. Xenopus oocyte germinal-vesicle breakdown induced by [Val12]Ras is inhibited by a cytosol-localized Ras mutant. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6630–6634. doi: 10.1073/pnas.86.17.6630. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hancock J. F., Magee A. I., Childs J. E., Marshall C. J. All ras proteins are polyisoprenylated but only some are palmitoylated. Cell. 1989 Jun 30;57(7):1167–1177. doi: 10.1016/0092-8674(89)90054-8. [DOI] [PubMed] [Google Scholar]
  19. Hurley J. B., Simon M. I., Teplow D. B., Robishaw J. D., Gilman A. G. Homologies between signal transducing G proteins and ras gene products. Science. 1984 Nov 16;226(4676):860–862. doi: 10.1126/science.6436980. [DOI] [PubMed] [Google Scholar]
  20. Korn L. J., Siebel C. W., McCormick F., Roth R. A. Ras p21 as a potential mediator of insulin action in Xenopus oocytes. Science. 1987 May 15;236(4803):840–843. doi: 10.1126/science.3554510. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Kubota H. Y., Yoshimoto Y., Yoneda M., Hiramoto Y. Free calcium wave upon activation in Xenopus eggs. Dev Biol. 1987 Jan;119(1):129–136. doi: 10.1016/0012-1606(87)90214-4. [DOI] [PubMed] [Google Scholar]
  23. Lacal J. C. Diacylglycerol production in Xenopus laevis oocytes after microinjection of p21ras proteins is a consequence of activation of phosphatidylcholine metabolism. Mol Cell Biol. 1990 Jan;10(1):333–340. doi: 10.1128/mcb.10.1.333. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lacal J. C., de la Peña P., Moscat J., Garcia-Barreno P., Anderson P. S., Aaronson S. A. Rapid stimulation of diacylglycerol production in Xenopus oocytes by microinjection of H-ras p21. Science. 1987 Oct 23;238(4826):533–536. doi: 10.1126/science.2821623. [DOI] [PubMed] [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. Lucas A. M., Thody A. J., Shuster S. Role of protein kinase C in the pigment cell of the lizard (Anolis carolinensis). J Endocrinol. 1987 Feb;112(2):283–287. doi: 10.1677/joe.0.1120283. [DOI] [PubMed] [Google Scholar]
  27. Melton D. A., Krieg P. A., Rebagliati M. R., Maniatis T., Zinn K., Green M. R. Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. Nucleic Acids Res. 1984 Sep 25;12(18):7035–7056. doi: 10.1093/nar/12.18.7035. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Parker I., Miledi R. Changes in intracellular calcium and in membrane currents evoked by injection of inositol trisphosphate into Xenopus oocytes. Proc R Soc Lond B Biol Sci. 1986 Aug 22;228(1252):307–315. doi: 10.1098/rspb.1986.0057. [DOI] [PubMed] [Google Scholar]
  29. Price B. D., Morris J. D., Marshall C. J., Hall A. Stimulation of phosphatidylcholine hydrolysis, diacylglycerol release, and arachidonic acid production by oncogenic ras is a consequence of protein kinase C activation. J Biol Chem. 1989 Oct 5;264(28):16638–16643. [PubMed] [Google Scholar]
  30. Reynhout J. K., Taddei C., Smith L. D., LaMarca M. J. Response of large oocytes of Xenopus laevis to progesterone in vitro in relation to oocyte size and time after previous HCG-induced ovulation. Dev Biol. 1975 Jun;44(2):375–379. doi: 10.1016/0012-1606(75)90408-x. [DOI] [PubMed] [Google Scholar]
  31. Sadler S. E., Maller J. L. A similar pool of cyclic AMP phosphodiesterase in Xenopus oocytes is stimulated by insulin, insulin-like growth factor 1, and [Val12,Thr59]Ha-ras protein. J Biol Chem. 1989 Jan 15;264(2):856–861. [PubMed] [Google Scholar]
  32. 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]
  33. Smith L. D. The induction of oocyte maturation: transmembrane signaling events and regulation of the cell cycle. Development. 1989 Dec;107(4):685–699. doi: 10.1242/dev.107.4.685. [DOI] [PubMed] [Google Scholar]
  34. Sweet R. W., Yokoyama S., Kamata T., Feramisco J. R., Rosenberg M., Gross M. The product of ras is a GTPase and the T24 oncogenic mutant is deficient in this activity. Nature. 1984 Sep 20;311(5983):273–275. doi: 10.1038/311273a0. [DOI] [PubMed] [Google Scholar]
  35. Taylor M. A., Johnson A. D., Smith L. D. Growing Xenopus oocytes have spare translational capacity. Proc Natl Acad Sci U S A. 1985 Oct;82(19):6586–6589. doi: 10.1073/pnas.82.19.6586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Taylor M. A., Smith L. D. Induction of maturation in small Xenopus laevis oocytes. Dev Biol. 1987 May;121(1):111–118. doi: 10.1016/0012-1606(87)90144-8. [DOI] [PubMed] [Google Scholar]
  37. Taylor M. A., Smith L. D. Quantitative changes in protein synthesis during oogenesis in Xenopus laevis. Dev Biol. 1985 Jul;110(1):230–237. doi: 10.1016/0012-1606(85)90079-x. [DOI] [PubMed] [Google Scholar]
  38. Wallace R. A., Jared D. W., Dumont J. N., Sega M. W. Protein incorporation by isolated amphibian oocytes. 3. Optimum incubation conditions. J Exp Zool. 1973 Jun;184(3):321–333. doi: 10.1002/jez.1401840305. [DOI] [PubMed] [Google Scholar]
  39. Willumsen B. M., Norris K., Papageorge A. G., Hubbert N. L., Lowy D. R. Harvey murine sarcoma virus p21 ras protein: biological and biochemical significance of the cysteine nearest the carboxy terminus. EMBO J. 1984 Nov;3(11):2581–2585. doi: 10.1002/j.1460-2075.1984.tb02177.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Wolfman A., Moscucci A., Macara I. G. Evidence for multiple, ras-like, guanine nucleotide-binding proteins in Swiss 3T3 plasma membranes. Stimulation of GTPase activity by cytosolic factors. J Biol Chem. 1989 Jun 25;264(18):10820–10827. [PubMed] [Google Scholar]

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