Localization of NuMA protein isoforms in the nuclear matrix of mammalian cells

Cell Motil Cytoskeleton. 1994;29(2):167-76. doi: 10.1002/cm.970290208.

Abstract

Using a monoclonal antibody 2D3 generated against a kinetochore-enriched human chromosome preparation, we identified a high molecular mass protein with nuclear staining in interphase and polar staining of the pericentriolar region in the mitotic spindle. Initially termed centrophilin, this protein associates with the minus-ends of spindle microtubules (MT) and appears to be important in spindle organization [Tousson et al., 1991: J. Cell Biol. 112:427-440]. Comparison of a partial cDNA sequence obtained for centrophilin with the full length cDNA sequence of nuclear mitotic apparatus protein (NuMA) [Compton et al., 1992: J. Cell Biol. 116:1395-1408; Yang et al., 1992: J. Cell Biol. 116:1303-1317] has indicated that NuMA and centrophilin are the same protein. Using a polyclonal NuMA antibody, we have provided further evidence that NuMA exists as isoforms as shown by peptide mapping and immunoblots. Sequential fractionation experiments along with immunofluorescence, immunoblotting, and EM immunogold labeling have demonstrated that NuMA isoforms are novel components of nuclear core filaments. Thus, NuMA, a long coiled-coil protein, appears to have dual functions in interphase and mitosis during the cell cycle. In interphase, NuMA likely plays a structural role in the nucleoskeleton that may be important in nuclear organization and functions, whereas in mitosis, NuMA appears to be associated with spindle MT organization and chromosome positioning.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antigens, Nuclear
  • Carcinoma, Squamous Cell
  • Female
  • HeLa Cells
  • Humans
  • Immunoblotting
  • Immunohistochemistry
  • Interphase
  • Isomerism
  • Microscopy, Fluorescence
  • Microscopy, Immunoelectron
  • Microtubules / chemistry
  • Mitosis
  • Nuclear Matrix / chemistry*
  • Nuclear Matrix / ultrastructure
  • Nuclear Matrix-Associated Proteins
  • Nuclear Proteins / analysis*
  • Nuclear Proteins / chemistry
  • Nuclear Proteins / physiology
  • Peptide Mapping
  • Tumor Cells, Cultured
  • Uterine Cervical Neoplasms

Substances

  • Antigens, Nuclear
  • NUMA1 protein, human
  • Nuclear Matrix-Associated Proteins
  • Nuclear Proteins