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Kinetochore structure, duplication, and distribution in mammalian cells: analysis by human autoantibodies from scleroderma patients

Academic Article
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Overview

authors

  • Brenner, S.
  • Pepper, D.
  • Berns, M. W.
  • Tan, Eng
  • Brinkley, B. R.

publication date

  • 1981

journal

  • Journal of Cell Biology  Journal

abstract

  • The specificity of the staining of CREST scleroderma patient serum was investigated by immunofluorescence and immunoelectron microscopy. The serum was found to stain the centromere region of mitotic chromosomes in many mammalian cell types by immunofluorescence. It also localized discrete spots in interphase nuclei which we have termed "presumptive kinetochores." The number of presumptive kinetochores per cell corresponds to the chromosome number in the cell lines observed. Use of the immunoperoxidase technique to localize the antisera on PtK2 cells at the electron microscopic level revealed the specificity of the sera for the trilaminar kinetochore disks on metaphase and anaphase chromosomes. Presumptive kinetochores in the interphase nuclei were also visible in the electron microscope as randomly arranged, darkly stained spheres averaging 0.22 micrometers in diameter. Preabsorption of the antisera was attended using microtubule protein, purified tubulin, actin, and microtubule-associated proteins. None of these proteins diminished the immunofluorescence staining of the sera, indicating that the antibody-specific antigen(s) is a previously unrecognized component of the kinetochore region. In some interphase cells observed by both immunofluorescence and immunoelectron microscopy, the presumptive kinetochores appeared as double rather than single spots. Analysis of results obtained using a microspectrophotometer to quantify DNA in individual cells double stained with scleroderma serum and the DNA fluorescent dye, propidium iodide, led to the conclusion that the presumptive kinetochores duplicate in G2 of the cell cycle.

subject areas

  • Animals
  • Autoantibodies
  • Cell Cycle
  • Centromere
  • Chromatin
  • Chromosomes
  • Demecolcine
  • Humans
  • Microtubules
  • Mitosis
  • Scleroderma, Systemic
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Identity

PubMed Central ID

  • PMC2111947

International Standard Serial Number (ISSN)

  • 0021-9525

Digital Object Identifier (DOI)

  • 10.1083/jcb.91.1.95

PubMed ID

  • 7298727
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Additional Document Info

start page

  • 95

end page

  • 102

volume

  • 91

issue

  • 1

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