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. 1981 Feb 1;88(2):410–421. doi: 10.1083/jcb.88.2.410

Involvement of contractile proteins in the changes in consistency of oocyte nucleoplasm of the newt Pleurodeles waltlii

PMCID: PMC2111746  PMID: 6894149

Abstract

In the present work, we show that actin is present in considerable quantities in the oocyte nucleus of the newt Pleurodeles waltlii. The nuclear sap, extracted in saline buffer containing Ca++, is fluid. DNAase I inhibition assays have shown that 90% of actin is under a globular state in such conditions. Chelation of Ca++ by EGTA leads to the formation of a nuclear gel composed of individual microfilaments. This nuclear gel contains approximately 50% of total nuclear actin in a filamentous form. Phalloidin, a drug known to stabilize F-actin, induces the formation of a network of actin cables in the nuclei. This network contains nearly 100% of total nuclear actin in the filamentous form. The observation of the cables in the electron microscope shows that they are made of tightly associated microfilaments to which RNP- like particles are bound. The actin antibodies stain the cables and the particles by the indirect immunoperoxidase technique; myosin antibodies mainly stain the particles. The formation of the phalloidin-induced network seems to require the presence of Ca++, Mg++, and ATP. We propose a scheme for the regulation of the supramolecular forms of actin in oocyte nuclei in which a delicate equilibrium seems to exist between globular actin, microfilaments, and actin cables. This equilibrium would be controlled by the concentration of Ca++, ATP, and various actin-associated proteins.

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Selected References

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  1. Blikstad I., Markey F., Carlsson L., Persson T., Lindberg U. Selective assay of monomeric and filamentous actin in cell extracts, using inhibition of deoxyribonuclease I. Cell. 1978 Nov;15(3):935–943. doi: 10.1016/0092-8674(78)90277-5. [DOI] [PubMed] [Google Scholar]
  2. Brunel C., Lelay M. N. Two-dimensional analysis of proteins associated with heterogenous nuclear RNA in various animal cell lines. Eur J Biochem. 1979 Sep;99(2):273–283. doi: 10.1111/j.1432-1033.1979.tb13254.x. [DOI] [PubMed] [Google Scholar]
  3. CALLAN H. G. THE NATURE OF LAMPBRUSH CHROMOSOMES. Int Rev Cytol. 1963;15:1–34. doi: 10.1016/s0074-7696(08)61114-6. [DOI] [PubMed] [Google Scholar]
  4. Callan H. G. Chromosomes and nucleoli of the axolotl, Ambystoma mexicanum. J Cell Sci. 1966 Mar;1(1):85–108. doi: 10.1242/jcs.1.1.85. [DOI] [PubMed] [Google Scholar]
  5. Clark T. G., Merriam R. W. Diffusible and bound actin nuclei of Xenopus laevis oocytes. Cell. 1977 Dec;12(4):883–891. doi: 10.1016/0092-8674(77)90152-0. [DOI] [PubMed] [Google Scholar]
  6. Clark T. G., Rosenbaum J. L. An actin filament matrix in hand-isolated nuclei of X. laevis oocytes. Cell. 1979 Dec;18(4):1101–1108. doi: 10.1016/0092-8674(79)90223-x. [DOI] [PubMed] [Google Scholar]
  7. Comings D. E., Harris D. C. Nuclear proteins. I. Electrophoretic comparison of mouse nucleoli, heterochromatin, euchromatin and contractile proteins. Exp Cell Res. 1975 Nov;96(1):161–179. doi: 10.1016/s0014-4827(75)80049-8. [DOI] [PubMed] [Google Scholar]
  8. Cuatrecasas P., Wilchek M., Anfinsen C. B. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968 Oct;61(2):636–643. doi: 10.1073/pnas.61.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Douvas A. S., Harrington C. A., Bonner J. Major nonhistone proteins of rat liver chromatin: preliminary identification of myosin, actin, tubulin, and tropomyosin. Proc Natl Acad Sci U S A. 1975 Oct;72(10):3902–3906. doi: 10.1073/pnas.72.10.3902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gabbiani G., Ryan G. B., Lamelin J. P., Vassalli P., Majno G., Bouvier C. A., Cruchaud A., Lüscher E. F. Human smooth muscle autoantibody. Its identification as antiactin antibody and a study of its binding to "nonmuscular" cells. Am J Pathol. 1973 Sep;72(3):473–488. [PMC free article] [PubMed] [Google Scholar]
  11. Goldstein L., Rubin R., Ko C. The presence of actin in nuclei: a critical appraisal. Cell. 1977 Nov;12(3):601–608. doi: 10.1016/0092-8674(77)90260-4. [DOI] [PubMed] [Google Scholar]
  12. Goody R. S., Holmes K. C., Mannherz H. G., Leigh J. B., Rosenbaum G. Cross-bridge conformation as revealed by x-ray diffraction studies on insect flight muscles with ATP analogues. Biophys J. 1975 Jul;15(7):687–705. doi: 10.1016/S0006-3495(75)85848-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gordon W. E., 3rd, Bushnell A., Burridge K. Characterization of the intermediate (10 nm) filaments of cultured cells using an autoimmune rabbit antiserum. Cell. 1978 Feb;13(2):249–261. doi: 10.1016/0092-8674(78)90194-0. [DOI] [PubMed] [Google Scholar]
  14. Graham R. C., Jr, Karnovsky M. J. The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique. J Histochem Cytochem. 1966 Apr;14(4):291–302. doi: 10.1177/14.4.291. [DOI] [PubMed] [Google Scholar]
  15. Hartwig J. H., Stossel T. P. Isolation and properties of actin, myosin, and a new actinbinding protein in rabbit alveolar macrophages. J Biol Chem. 1975 Jul 25;250(14):5696–5705. [PubMed] [Google Scholar]
  16. Hellewell S. B., Taylor D. L. The contractile basis of ameboid movement. VI. The solation-contraction coupling hypothesis. J Cell Biol. 1979 Dec;83(3):633–648. doi: 10.1083/jcb.83.3.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Herman I. M., Pollard T. D. Actin localization in fixed dividing cells stained with fluorescent heavy meromyosin. Exp Cell Res. 1978 Jun;114(1):15–25. doi: 10.1016/0014-4827(78)90030-7. [DOI] [PubMed] [Google Scholar]
  18. Hitchcock S. E. Regulation of motility in nonmuscle cells. J Cell Biol. 1977 Jul;74(1):1–15. doi: 10.1083/jcb.74.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jockusch B. M., Kelley K. H., Meyer R. K., Burger M. M. An efficient method to produce specific anti-actin. Histochemistry. 1978 Apr 4;55(3):177–184. doi: 10.1007/BF00495757. [DOI] [PubMed] [Google Scholar]
  20. Karsenti E., Guilbert B., Bornens M., Avrameas S. Antibodies to tubulin in normal nonimmunized animals. Proc Natl Acad Sci U S A. 1977 Sep;74(9):3997–4001. doi: 10.1073/pnas.74.9.3997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Karsenti E., Guilbert B., Bornens M., Avrameas S., Whalen R., Pantaloni D. Detection of tubulin and actin in various cell lines by an immunoperoxidase technique. J Histochem Cytochem. 1978 Nov;26(11):934–947. doi: 10.1177/26.11.363933. [DOI] [PubMed] [Google Scholar]
  22. Lengsfeld A. M., Löw I., Wieland T., Dancker P., Hasselbach W. Interaction of phalloidin with actin. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2803–2807. doi: 10.1073/pnas.71.7.2803. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lestourgeon W. M., Forer A., Yang Y. Z., Bertram J. S., Pusch H. P. Contractile proteins. Major components of nuclear and chromosome non-histone proteins. Biochim Biophys Acta. 1975 Feb 27;379(2):529–552. [PubMed] [Google Scholar]
  24. MOMMAERTS W. F. H. M., PARRISH R. G. Studies on myosin. I. Preparation and criteria of purity. J Biol Chem. 1951 Feb;188(2):545–552. [PubMed] [Google Scholar]
  25. Malcolm D. B., Sommerville J. The structure of chromosome-derived ribonucleoprotein in oocytes of Triturus cristatus carnifex (Laurenti). Chromosoma. 1974;48(2):137–158. doi: 10.1007/BF00283960. [DOI] [PubMed] [Google Scholar]
  26. Maundrell K., Scherrer K. Characterization of pre-messenger-RNA-containing nuclear ribonucleoprotein particles from avian erythroblasts. Eur J Biochem. 1979 Sep;99(2):225–238. doi: 10.1111/j.1432-1033.1979.tb13249.x. [DOI] [PubMed] [Google Scholar]
  27. Mott M. R., Callen H. G. An electron-microscope study of the lampbrush chromosomes of the newt Triturus cristatus. J Cell Sci. 1975 Mar;17(3):241–261. doi: 10.1242/jcs.17.3.241. [DOI] [PubMed] [Google Scholar]
  28. Paulin D., Nicolas J. F., Jacquet M., Jakob H., Gros F., Jacob F. Comparative protein patterns in chromatins from mouse teratocarcinoma cells. Exp Cell Res. 1976 Oct 1;102(1):169–178. doi: 10.1016/0014-4827(76)90312-8. [DOI] [PubMed] [Google Scholar]
  29. Pollard T. D., Thomas S. M., Niederman R. Human platelet myosin. I. Purification by a rapid method applicable to other nonmuscle cells. Anal Biochem. 1974 Jul;60(1):258–266. doi: 10.1016/0003-2697(74)90152-3. [DOI] [PubMed] [Google Scholar]
  30. Rubin R. W., Goldstein L., Ko C. Differences between nucleus and cytoplasm in the degree of actin polymerization. J Cell Biol. 1978 Jun;77(3):698–701. doi: 10.1083/jcb.77.3.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rungger D., Rungger-Brändle E., Chaponnier C., Gabbiani G. Intranuclear injection of anti-actin antibodies into Xenopus oocytes blocks chromosome condensation. Nature. 1979 Nov 15;282(5736):320–321. doi: 10.1038/282320a0. [DOI] [PubMed] [Google Scholar]

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