Chromosoma

, Volume 113, Issue 6, pp 316–323 | Cite as

Centromeric protein bodies on avian lampbrush chromosomes contain a protein detectable with an antibody against DNA topoisomerase II

  • Alla Krasikova
  • Tatiana Kulikova
  • Alsu Saifitdinova
  • Svetlana Derjusheva
  • Elena Gaginskaya
Research Article

Abstract

In the oocyte nuclei (germinal vesicle or GV) of a variety of avian species, prominent spherical entities termed protein bodies (PBs) arise at the centromeric regions of the lampbrush chromosomes (LBCs). In spite of the obvious protein nature of PBs, nothing is known about their composition. We show that an antibody against DNA topoisomerase II (topo II), the DNA unwinding enzyme, recognizes PBs from chaffinch and pigeon oocytes. In later chaffinch oocytes, the PBs fuse to form a karyosphere, which is also labeled by the anti-topo II antibody. Furthermore, we show that proteins characteristic of Cajal bodies and B-snurposomes are not found in PBs, despite morphological similarities among these structures. Using immunoelectron microscopy and immunofluorescent laser scanning microscopy we demonstrated that topo II localizes predominantly in the dense material of PBs. Two antigens of ∼170 kDa (which corresponds to topo II) and ∼100 kDa were revealed with the antibody against topo II on immunoblots of avian GV proteins. We propose that the smaller protein results from oocyte specific topo II cleavage, since it was not detected in nuclei from testis cells. This represents the first report of a defined protein in the centromeric PBs on avian LBCs.

References

  1. Adachi Y, Kas E, Laemmli UK (1989) Preferential, cooperative binding of DNA topoisomerase II to scaffold-associated regions. EMBO J 8:3997–4006PubMedGoogle Scholar
  2. Andersen CL, Wandall A, Kjeldsen E, Mielke C, Koch J (2002) Active, but not inactive, human centromeres display topoisomerase II activity in vivo. Chromosome Res 10:305–312CrossRefPubMedGoogle Scholar
  3. Andrade LEC, Tan EM, Chan EKL (1993) Immunocytochemical analysis of the coiled body in the cell cycle and during cell proliferation. Proc Natl Acad Sci USA 90:1947–1951PubMedGoogle Scholar
  4. Berezney R, Coffey DS (1974) Identification of nuclear protein matrix. Biochem Biophys Res Commun 60:1410–1417PubMedGoogle Scholar
  5. Berrios M, Osheroff N, Fisher PA (1985) In situ localization of DNA topoisomerase II, a major polypeptide component of the Drosophila nuclear matrix fraction. Proc Natl Acad Sci USA 82:4142–4146PubMedGoogle Scholar
  6. Callan HG (1986) Lampbrush chromosomes. Springer, Berlin Heidelberg New YorkGoogle Scholar
  7. Chang C-J, Goulding S, Earnshaw WC, Carmena M (2003) RNAi analysis reveals an unexpected role for topoisomerase II in chromosome arm congression to a metaphase plate. J Cell Sci 116:4715–4726CrossRefPubMedGoogle Scholar
  8. Cobb J, Miyaike M, Kikuchi A, Handel MA (1999) Meiotic events at the centromeric heterochromatin: histone H3 phosphorylation, topoisomerase IIα localization and chromosome condensation. Chromosoma 108:412–425CrossRefPubMedGoogle Scholar
  9. Doyle O, Corden JL, Murphy C, Gall JG (2002) The distribution of RNA polymerase II largest subunit (RPB1) in the Xenopus germinal vesicle. J Struct Biol 140:154–166CrossRefPubMedGoogle Scholar
  10. Earnshaw WC, Heck MMS (1985) Localization of topoisomerase II in mitotic chromosomes. J Cell Biol 100:1716–1725CrossRefPubMedGoogle Scholar
  11. Earnshaw WC, Halligan B, Cooke CA, Heck MMS, Liu LF (1985) Topoisomerase II is a structural component of mitotic chromosome scaffolds. J Cell Biol 100:1706–1715CrossRefPubMedGoogle Scholar
  12. Fisher D, Hock R, Sheer U (1993) DNA topoisomerase II is not detectable on lampbrush chromosomes but enriched in the amplified nucleoli of Xenopus oocytes. Exp Cell Res 209:255–260CrossRefPubMedGoogle Scholar
  13. Fu X-D, Maniatis T (1990) Factor required for mammalin spliceosome assambly is localized to discrete regions in the nucleus. Nature 343:437–441CrossRefPubMedGoogle Scholar
  14. Gaginskaya ER (1972) Nuclear structures in oocytes of adult birds. II. Protein bodies and the karyosphere. Tsitologiia 14:568–578PubMedGoogle Scholar
  15. Gaginskaya ER, Gruzova MN (1969) Characteristics of oogenesis in the finch. Tsitologiia 11:1241–1251PubMedGoogle Scholar
  16. Gaginskaya ER, Gruzova MN (1975) Detection of the amplified rDNA in ovarial cells of some insects and birds by hybridization in situ. Tsitologiia 17:1132–1137Google Scholar
  17. Gall JG (2000) Cajal bodies: the first 100 years. Annu Rev Cell Dev Biol 16:273–300CrossRefPubMedGoogle Scholar
  18. Gall JG (2003) The centennial of the Cajal body. Nat Rev Mol Cell Biol 4: 975–980CrossRefPubMedGoogle Scholar
  19. Gall JG, Callan HG (1989) The sphere organelle contains small nuclear ribonucleoproteins. Proc Natl Acad Sci USA 86:6635–6639PubMedGoogle Scholar
  20. Gall JG, Bellini M, Wu Z, Murphy C (1999) Assembly of the nuclear transcription and processing machinery: Cajal bodies (coiled bodies) and transcriptosomes. Mol Biol Cell 10:4385–4402PubMedGoogle Scholar
  21. Heck MMS, Hittelman WN, Earnshaw WC (1988) Differential expression of DNA topoisomerase I and II during the eukaryotic cell cycle. Proc Natl Acad Sci USA 85:1086–1090PubMedGoogle Scholar
  22. Hock R, Carl M, Lieb B, Gebauer D, Scheer U (1996) A monoclonal antibody against DNA topoisomerase II labels the axial granules of Pleurodeles lampbrush chromosomes. Chromosoma 104:358–366CrossRefPubMedGoogle Scholar
  23. Lerner EA, Lerner MR, Janeway CA, Steitz JA (1981) Monoclonal antibodies to nucleic acid-containing cellular constituents: probes for molecular biology and autoimmune disease. Proc Natl Acad Sci USA 78:2737–2741PubMedGoogle Scholar
  24. Mondal N, Parvin JD (2001) DNA topoisomerase IIα is required for RNA polymerase II transcription on chromatin templates. Nature 413:435–438CrossRefPubMedGoogle Scholar
  25. Morgan GT (2002) Lampbrush chromosomes and associated bodies: new insights into principles of nuclear structure and function. Chromosome Res 10:177–200CrossRefPubMedGoogle Scholar
  26. Morgan GT, Doyle O, Murphy C, Gall JG (2000) RNA polymerase II in Cajal bodies of amphibian oocytes. J Struct Biol 129:258–268CrossRefPubMedGoogle Scholar
  27. Newman GR, Hobot JA (1993) Resin microscopy and on-section immunocytochemistry. Springer, Berlin Heidelberg New YorkGoogle Scholar
  28. Niimi A, Suka N, Harata M, Kikuchi A, Mizuno S (2001) Co-localization of chicken DNA topoisomerase IIα, but not β, with sites of DNA replication and possible involvement of a C-terminal region of α through its binding to PCNA. Chromosoma 110:102–114PubMedGoogle Scholar
  29. Olson M, Dundr M, Szebeni A (2000) The nucleolus: an old factory with unexpected capabilities. Trends Cell Biol 10:189–196CrossRefPubMedGoogle Scholar
  30. Paliulis LV, Nicklas RB (2003) Topoisomerase II may be linked to the reduction of chromosome number in meiosis. BioEssays 25:309–312CrossRefPubMedGoogle Scholar
  31. Patturajan M, Schulte R, Sefton B, Berezney R, Vincent M, Bensaude O, Warren S, Corden J (1998) Growth-related changes in phosphorylation of yeast RNA polymerase II. J Biol Chem 273:4689–4694CrossRefPubMedGoogle Scholar
  32. Pollard KM, Lee DK, Casiano CA, Bluthner M, Johnston MM, Tan EM (1997) The autoimmunity-inducing xenobiotic mercury interacts with the autoantigen fibrillarin and modifies its molecular and antigenic properties. J Immunol 158:3521–3528PubMedGoogle Scholar
  33. Saifitdinova A, Derjusheva S, Krasikova A, Gaginskaya E (2003) Lampbrush chromosomes of the chaffinch (Fringilla coelebs L.). Chromosome Res 11:93–113CrossRefGoogle Scholar
  34. Salmena L, Lam V, McPherson JP, Goldenberg GJ (2001) Role of proteasomal degradation in the cell cycle-dependent regulation of DNA topoisomerase IIalpha expression. Biochem Pharmacol 61:795–802CrossRefPubMedGoogle Scholar
  35. Schmidt-Zachmann MS, Franke WW (1988) DNA cloning and amino acid sequence determination of a major constituent protein of mammalian nucleoli. Correspondence of the nucleoplasmin-related protein NO38 to mammalian protein B23. Chromosoma 96:417–426CrossRefPubMedGoogle Scholar
  36. Solovei I, Gaginskaya E, Hutchison N, Macgregor H (1993) Avian sex chromosomes in the lampbrush form: the ZW lampbrush bivalent from six species of bird. Chromosome Res 1:153–166PubMedGoogle Scholar
  37. Solovei IV, Gaginskaya ER, Macgregor HC (1994) The arrangement and transcription of telomere DNA sequences at the ends of lampbrush chromosomes of birds. Chromosome Res 2:460–470PubMedGoogle Scholar
  38. Solovei IV, Joffe BI, Gaginskaya ER, Macgregor HC (1996) Transcription on lampbrush chromosomes of a centromerically localized highly repeated DNA in pigeon (Columba) relates to sequence arrangement. Chromosome Res 4:588–603PubMedGoogle Scholar
  39. St Pierre J, Wright DJ, Rowe TC, Wright SJ (2002) DNA topoisomerase II distribution in mouse preimplantation embryos. Mol Reprod Dev 61:335–346CrossRefPubMedGoogle Scholar
  40. Swedlow JR, Hirano T (2003) The making of the mitotic chromosome: modern insights into classical questions. Mol Cell 11:557–569CrossRefPubMedGoogle Scholar
  41. Tsvetkov AG, Gaginskaya ER (1983) The nuclear matrix of oocytes of the chaffinch (Fringilla coelebs L.). Tsitologiia 25:649–654Google Scholar
  42. Wu Z, Murphy C, Callan HG, Gall JG (1991) Small nuclear ribonucleoproteins and heterogeneous nuclear ribonucleoproteins in the amphibian germinal vesicle: loops, spheres, and snurposomes. J Cell Biol 113:465–483CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Alla Krasikova
    • 1
  • Tatiana Kulikova
    • 1
  • Alsu Saifitdinova
    • 1
  • Svetlana Derjusheva
    • 1
  • Elena Gaginskaya
    • 1
  1. 1.Biological Research InstituteSaint-Petersburg State UniversitySaint PetersburgRussia

Personalised recommendations