Chromosoma

, Volume 105, Issue 1, pp 31–40 | Cite as

Postmetaphase nuclear formation: Loss of a chromosomal epitope coincident with apparent chromatid coalescence

  • D. L. Adams
  • L. D. Hodge
Article
  • 31 Downloads

Abstract

Previously, we have conceptualized mitotic nuclear formation following metaphase as a morphogenic process and have suggested that sets of chromatids, after separation from a metaphase plate, can be thought of as prenuclei. Such structures can be grouped temporally as either early or late prenuclei based on morphologic, morphometric and density characteristics. Sequential ordering of early prenuclei is of particular interest because it reveals that condensed chromatids coalesce with the resulting formation of a unique chambered structure. In this paper we describe data obtained with a newly raised monoclonal antibody (mAb-2) that initially recognizes an epitope(s) on metaphase chromosomes. Light and confocal fluorescent microscopy of early prenuclei reveal that the chromosomal epitope can no longer be detected about chromatids after their apparent coalescence. Immunoblot analysis of dispersed polypeptides of metaphase plates and early prenuclei indicates that the major protein antigens recognized by mAb-2 have apparent molecular masses of approximately 106000 and 80500 and that each is likely composed of multiple charge isomers. A dual fluorescent analysis using mAb-2 and high-titer anti-lamin B serum provides additional evidence that chromatid coalescence is a separate, early event that precedes nuclear lamina formation.

Keywords

Nuclear Pore Complex Metaphase Plate Nuclear Periphery Phase Plate Central Chamber 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Baserga BJ, Yang XD, Steitz JA (1991) An intact Box C sequence in the U3 snRNA is required for binding of fibrillarin, the protein common to the major family of nucleolar snRNPs. EMBO J 10:2645–2651PubMedGoogle Scholar
  2. Chaly N, Bladon T, Setterfield G, Little JE, Kaplan JG, Brown DL (1984) Changes in distribution of nuclear matrix during the cell cycle. J Cell Biol 99:661–671PubMedCrossRefGoogle Scholar
  3. Forbes DJ (1992) Structure and function of the nuclear pore complex. Annu Rev Cell Biol 8:495–527PubMedCrossRefGoogle Scholar
  4. Gerace L, Blobel G (1980) The nuclear envelope lamina is reversibly depolymerized during mitosis. Cell 19:277–287PubMedCrossRefGoogle Scholar
  5. Hall CT, Hansen PA (1962) Chelated azo dyes used as counterstains in the fluorescent antibody techniques. Zentralbl Bakteriol Mikrobiol Hyg [B], 184:548–555Google Scholar
  6. Harlow E, Lane D (1988). Antibodies: a laboratory manual. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NYGoogle Scholar
  7. Henry SM, Hodge LD (1983) Nuclear matrix: a cell cycle dependent site of increased intranuclear protein phosphorylation. Eur J Biochem 133:194–208CrossRefGoogle Scholar
  8. Hodge LD, Mancini P, Davis FM, Heywood P (1977) Nuclear matrix of HeLa S3 cells: Polypeptide composition during adenovirus infection and in phases of the cell cycle. J Cell Biol 72:194–208PubMedCrossRefGoogle Scholar
  9. Hodge LD, Martinez SE, Allsbrook WC, Pantazis CG, Welter DA (1990) Intermediate structures in nuclear morphogenesis following metaphase from HeLa S3 cells can be isolated and temporally grouped. Chromosoma 99:169–182PubMedCrossRefGoogle Scholar
  10. Kamei H (1990) Location of nuclear antigens recognized by DSB 389 MAB, a monoclonal antibody against desmin, observed by confocal laser scanning fluorescence microscopy. Cell Biol Inf Rep 14:727–736CrossRefGoogle Scholar
  11. McKeon FD, Tuffanelli DL, Kobayashi S, Kirschner MV (1984) The redistribution of a conserved nuclear envelope protein during the cell cycle suggests a pathway for chromosome condensation, Cell 36:83–92PubMedCrossRefGoogle Scholar
  12. Ottaviano Y, Gerace L (1985) Phosphorylations of the nuclear lamins during interphase and mitosis. J Biol Chem 260:624–632PubMedGoogle Scholar
  13. Snow CM, Senior A, Gerace L (1987) Monoclonal antibodies identifying a group of nuclear pore complex of glycoproteins. J Cell Biol 104:1143–1156PubMedCrossRefGoogle Scholar
  14. Welter DA, Black DA, Hodge LD (1985) Nuclear reformation following metaphase in HeLaS3 cells: three-dimensional visualization of chromatid rearrangements. Chromosoma 93:57–69PubMedCrossRefGoogle Scholar
  15. Welter DA, Black DA, Hodge LD (1986) Chromatid behavior in late mitosis: a scanning electron microscopy analysis of mammalian cell lines with various chromosome numbers. Scanning Electron Microsc IV:1371–1379Google Scholar
  16. Yasuda Y, Maul JJ (1990) A nucleolar auto-antigen is part of a major chromosomal surface component. Chromosoma 99:152–160PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • D. L. Adams
    • 1
  • L. D. Hodge
    • 1
  1. 1.Department of Cellular Biology and AnatomyMedical College of GeorgiaAugustaUSA

Personalised recommendations