Skip to main content
SpringerLink
Log in

Association of actin with DNA and nuclear matrix from Guerin ascites tumour cells

  • Original Article
  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

The protein composition of nuclear matrices containing different amount of DNA was examined. It was found that, in matrices containing 2% to 80% of total DNA, the quantity of DNA-bound proteins remains relatively constant varying from 10% to 15% of total nuclear proteins. Electrophoretic patterns do not differ substantially, but autoradiograms with in vitro 125I labelled proteins show quantitative variations in the actin content. Application of radioimmunoassay (RIA) enabled to determine the exact content of actin in GAT nuclei and nuclear matrices – 5 μg/ml in nuclei, of which 50% are bound to DNA and 3001o being a component of the protein part of the nuclear matrix. These results are supported by electron microscopic data, where immunogold technique was performed on thin sections and spread material. The applied methods suggest that part of the nuclear actin is tightly bound (resistant to 2 M NaCI) to DNA and represents a component of the internal nuclear matrix.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Jockush BM, Becker M, Hindennach I, Jockush H: Slime mould actin: homology to vertebrate actin and presence in the nucleus. Exp Cell Res 89:241–246, 1974

    Google Scholar 

  2. Douvas AS, Harrington CA, Bonner J: Major nonhistone proteins of rat liver chromatin: preliminary identification of myosin, actin, tubulin and tropomyosin. Proc Natl Acad Sci USA 72:3902–3906, 1975

    Google Scholar 

  3. Fukui Y, Katsumaru H: Dynamics of nuclear actin bundle induction by dymethyl sulfoxide and factors affecting its development. J Cell Biol 84:131–140, 1980

    Google Scholar 

  4. Gounon P, Karsenti E: Involvement of contractile proteins in the changes in consistency of oocyte nucleoplasm of the newt Pleurodeles waltlii. J Cell Biol 88:410–421, 1981

    Google Scholar 

  5. LeStourgeon WM, Forer A, Yang YZ, Bertram JS, Rusch H: Contractile proteins. Major components of nuclear and chromoome non-histone proteins. Biochem Biophys Acta 379:529–552, 1975

    Google Scholar 

  6. LeStourgeon WM: The occurance of contractile proteins in nuclei and their possible functions. In: H Busch (ed) The Cell Nucleus. Academic Press, New York 1978, pp 305–326

    Google Scholar 

  7. Egly JM, Miyamoto NG, Moncollin V, Chambon P: Is actin a transcription initiation factor for RNA polymerase β? EMBO J 3:2363–2371, 1984

    Google Scholar 

  8. Scheer U, Hinssen H, Franke WW, Jockush BM: Microinjection of actin-binding proteins and actin antibodies demonstrates involvement of nuclear actin in transcription of lampbrush chromosomes. Cell 39:111–122, 1984

    Google Scholar 

  9. Nakayasu H, Mori H, Ueda K: Association of small nuclear RNA-protein complex with the nuclear matrix from bovine lymphocytes. Cell Struct Funct 7:253–262, 1982

    Google Scholar 

  10. Nakayasu H, Ueda K: Small nuclear RNA-protein complex anchors on the actin filaments in bovine lymphocyte nuclear matrix. Cell Struct Funct 9:317–325, 1984

    Google Scholar 

  11. Nakayasu H, Ueda K: Association of rapidly-labeled RNAs with actin filaments in nuclear matrix from mouse L5178Y cells. Exp Cell Res 160:319–329, 1985

    Google Scholar 

  12. Rungger D, Rungger-Brandle E, Chaponnier C, Gabbiani G: Intranuclear injection of anti-actin antibodies into Xenopus oocytes blocks chromosome condensation. Nature 282:320–321, 1979

    Google Scholar 

  13. Karsenti E, Gounon P: Lampbrush chromosomes: are they ATP-dependent contractile structures? Biol Cell 34:91–98, 1979

    Google Scholar 

  14. Forer A: Does actin produce the force that moves a chromosome to the pole during anaphase? Can J Biochem Cell Biol 63:585–598, 1985

    Google Scholar 

  15. Bremer JW, Busch H, Yeoman CL: Evidence for a species of nuclear actin distinct from cytoplasmic and muscle actins. Biochemistry 20:2013–2017, 1981

    Google Scholar 

  16. Armbruster BL, Wunderli H, Turner BM, Raska I, Kellenberger E: Immunocytochemical localization of cytoskeletal proteins and histone 2B in isolated membrane-depleted nuclei, metaphase chromatin, and whole Chinese hamster ovary cells. J Histochem Cytochem 31:1385–1393, 1983

    Google Scholar 

  17. Nakayasu H, Ueda K: Association of actin with the nuclear matrix from bovine lymphocytes. Exp Cell Res 143:55–62, 1983

    Google Scholar 

  18. Nakayasu H, Ueda K: Ultrastructural localization of actin in nuclear matrices from mouse leukemia L5178Y cells. Cell Struct Funct 10:305–309, 1985

    Google Scholar 

  19. Hancock R: Topological organization of interphase DNA: the nuclear matrix and other skeletal structures. Biol Cell 46:105–122, 1982

    Google Scholar 

  20. Pienta KJ, Coffey DS: A structural analysis of the role of the nuclear matrix and DNA loops in the organization of the nucleus and chromosomes. J Cell Sci Suppl 1:123–135, 1984

    Google Scholar 

  21. Nelson WG, Pienta KJ, Barrack ER, Coffey DS: The role of the nuclear matrix in the organization and function of DNA. Ann Rev Biophys Chem 15:457–475, 1986

    Google Scholar 

  22. Krachmarov C, Iovcheva C, Hancock R, Dessev G: Association of DNA with the nuclear lamina in Ehrlich ascites tumour cells. J Cell Biochem 31:59–74, 1986

    Google Scholar 

  23. Hubert J, Bourgeois CA: The nuclear skeleton and the spatial arrangement of chromosomes in the interphase nucleus of vertebrate somatic cells. Hum Genet 74:1–15, 1986]

    Google Scholar 

  24. Kaufmann SH, Fields AP, Shaper JH: The nuclear matrix: current concepts and unanswered questions. Meth Achiev Exp Pathol 12:141–171, 1986

    Google Scholar 

  25. Turner BM, Franchi L: Identification of protein antigens associated with the nuclear matrix and with clusters of interchromatin granules in both interphase and mitotic cells. J Cell Sci 87:269–282, 1987

    Google Scholar 

  26. Hancock R, Hughes ME: Organization of DNA in the interphase nucleus. Biol Cell 44:201–212, 1982

    Google Scholar 

  27. Traub P, Nelson WJ, Kuhn S, Vorgias CE: The interaction in vitro of the intermediate filament protein vimentin with naturally occuring RNAs and DNAs. J Biol Chem 258:1456–1466, 1983

    Google Scholar 

  28. Kuhn S, Vorgias CE, Traub P: Interaction in vitro of nonepithelial intermediate filament proteins with supercoiled plasmid DNA. J Cell Sci 87:543–554, 1987

    Google Scholar 

  29. Earnshaw WC, Heck MMS: Localization of topoisomerase II in mitotic chromosomes. J Cell Biol 100:1716–1725, 1985

    Google Scholar 

  30. Gasser SM, Laroche T, Falquet J, Boy de la Tour E, Laemmli UK: Metaphase chromosome structure: Involvement of topoisomerase II. J Mol Biol 188:613–629, 1986

    Google Scholar 

  31. Gross DS, Garrard WT: Poising chromatin for transcription. Trends Biochem Sci 12:293–297, 1987

    Google Scholar 

  32. Avramova Z, Tsanev R: Stable DNA-protein complexes in eueukaryotic chromatin. J Mol Biol 196: 437–440, 1987

    Google Scholar 

  33. Neuer B, Plagens U, Werner D: Phosphordiester bonds between polypeptides and chromosomal DNA. J Mol Biol 164:213–235, 1983

    Google Scholar 

  34. Franke WW, Denk H, Kalt R, Schmid E: Biochemical and immunological identification of cytokeratin proteins present in hepatocytes of mammalian liver tissue. Exp Cell Res 131:299–318, 1981

    Google Scholar 

  35. Uschewa A, Patriotis Ch, Avramova Z: An Hl-like protein from the sperm chromatin of Mytilus galloprovincialis. Cell Biol Int Rep 9:253–263, 1985

    Google Scholar 

  36. Charles R, De Graaf H, Mooraman AFM: Radioimmunochemical determination of carbamoyl-phosphat synthetase content of adult rat liver. Biochem Biophys Acta 629:36–49, 1980

    Google Scholar 

  37. Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685, 1970

    Google Scholar 

  38. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275, 1951

    Google Scholar 

  39. Burton K, Peterson GB: The quantitative distribution of pyrimidine nucleotides in calf thymus deoxyribonucleic acid. Biochem Biophys Acta 26:667–668, 1957

    Google Scholar 

  40. Coombs DH, Pearson GD: Filter binding assay for covalent DNA-protein complexes: Adenovirus DNA-terminal protein complex. Proc Natl Acad Sci USA 75:5291–5295, 1978

    Google Scholar 

  41. Miller OL, Bakken AH: Morphological studies on transcription. Acta Endocrynol Suppl 168:155–173, 1972

    Google Scholar 

  42. Frens G: Controlled nucleation of the regulation of particle size in monodisperse gold suspension. Nature 241:20–23, 1973

    Google Scholar 

  43. DeMey J, Moeremans M, Genens G, Nuydens R, DeBrabander M: High resolution light and electron microscopic localization of tubulin with the IGS (Immunogold Staining) method. Cell Biol Int Rep 5:889–900, 1981

    Google Scholar 

  44. Berezney R, Bucholtz LA: Dynamic association of replicating DNA fragments with the nuclear matrix of regenerating liver. Exp Cell Res 132:1–13, 1981

    Google Scholar 

  45. Small D, Nelkin B, Vogelstein B: The association of transcribed genes with the nuclear matrix of Drosophila cells during heat shock. Nucleic Acids Res 13:2413–2431, 1985

    Google Scholar 

  46. Comings DE, Wallack AS: DNA-binding properties of nuclear matrix proteins. J Cell Sci 34:233–246, 1978

    Google Scholar 

  47. McCready SJ, Akrigg A, Cook PR: Electron microscopy of intact nuclear DNA from human cells. J Cell Sci 39:53–62, 1979

    Google Scholar 

  48. Tsanev R, Avramova Z: Nonprotamine nucleoprotein ultrastructures in mature ram sperm nuclei. Eur J Cell Biol 24:139–145, 1981

    Google Scholar 

  49. Blickstad 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 15:935–943, 1978

    Google Scholar 

  50. Mannherz AG, Goody RS, Konrad M, Nowak E: The interaction of bovine pancreatic deoxyribonuclease I and skeletal muscle actin. Eur J Biochem 104:367–379, 1980

    Google Scholar 

  51. Kirov N, Djondjurov L, Tsanev R: Nuclear matrix and transcriptional activity of the mouse α-globin gene. J Mol Biol 180:601–614, 1984

    Google Scholar 

  52. Clark TG, Rosenbaum JL: An actin filament matrix in handisolated nuclei of Xenopus laevis oocytes. Cell 18:1101–1108, 1979

    Google Scholar 

  53. Jackson DA, McCready SJ, Cook PR: Replication and transcription depend on attachment of DNA to the nuclear cage. J Cell Sci Suppl 1:59–80, 1984

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institute of Molecular Biology, Bulgarian Academy of Sciences, 1113, Sofia, Bulgaria

    Nickola I. Valkov, Maja I. Ivanova, Anna A. Uscheva & Chavdar P. Krachmarov

Authors
  1. Nickola I. Valkov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maja I. Ivanova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anna A. Uscheva
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chavdar P. Krachmarov
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Valkov, N.I., Ivanova, M.I., Uscheva, A.A. et al. Association of actin with DNA and nuclear matrix from Guerin ascites tumour cells. Mol Cell Biochem 87, 47–56 (1989). https://doi.org/10.1007/BF00421082

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00421082

Key words

Search

Navigation