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Nuclear and mitotically enhanced epitope.

J C Hoffman1, J M Mullins

  • 1Department of Biology, Catholic University of America, Washington, DC 20064.

Cell Motility and the Cytoskeleton
|January 1, 1990
PubMed
Summary
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Monoclonal antibody C9 identifies a novel nuclear antigen present throughout the cell cycle, becoming exposed during mitosis. This antigen localizes to centrosomes, kinetochores, and the midbody during cell division.

Area of Science:

  • Cell Biology
  • Immunology
  • Molecular Biology

Background:

  • Mitosis involves dynamic changes in protein localization and function.
  • Understanding cell cycle regulation requires identifying key regulatory proteins.
  • Centrosomes, kinetochores, and the midbody are crucial structures during cell division.

Purpose of the Study:

  • To develop and characterize a novel monoclonal antibody (MAb) targeting mitotic-specific antigens.
  • To investigate the cell cycle-dependent localization and molecular identity of a newly identified nuclear antigen.

Main Methods:

  • Hybridoma production using salt-extracted proteins from taxol-stabilized microtubules.
  • Immunofluorescence microscopy for antigen localization.
  • Cell permeabilization and detergent extraction for antigen accessibility.

Related Experiment Videos

  • Immunoblot analysis for polypeptide identification across species.

    • Main Results:

    • Monoclonal antibody C9 recognized an epitope on an antigen localized to interphase centrosomes and nuclei.
    • The nuclear antigen's availability was cell cycle-dependent but present throughout the cycle upon permeabilization.
    • The antigen was exposed during prophase, released into the cytoplasm during nuclear envelope breakdown, and re-accumulated at daughter nuclei.
    • Mitotic cytoplasmic antigens localized to centrosomes, kinetochores, and the midbody.
    • Immunoblot analysis identified a ~250 kD polypeptide for the nuclear antigen and 107/117 kD and ~250 kD polypeptides for mitotic cytoplasmic antigens.

    Conclusions:

    • Monoclonal antibody C9 identifies a novel antigen with dynamic cell cycle-dependent localization.
    • This antigen plays a role in mitosis, associating with key structures like centrosomes and kinetochores.
    • The findings contribute to the understanding of nuclear and mitotic antigens and their roles in cell division.