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PMID: 1915296 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A 210 kDa nuclear matrix protein is a functional part of the mitotic spindle; a microinjection study using SPN monoclonal antibodies.

The EMBO journal ·Vol. 10 ·No. 11 ·1991-11-00 ·Pages 3351-62

Kallajoki M, Weber K, Osborn M

Abstract

Six monoclonal antibodies identify a 210 kDa polypeptide which shows a cell cycle specific redistribution from the nucleus to the mitotic spindle. In interphase cells this polypeptide was localized in the nucleus and behaved during differential cell extraction as a component of the nuclear matrix. It accumulated in the centrosome region at prophase, in the pole regions of the mitotic spindle at metaphase and in crescents at the poles in anaphase, and reassociated with the nuclei as they reformed in telophase. Due to its staining pattern we call the protein the Spindle Pole-Nucleus (SPN) antigen. The localization of SPN antigen during mitosis was dependent on the integrity of the spindle since treatment of cells with nocodazole resulted in the dispersal of SPN antigen into many small foci which acted as microtubule organizing centres when the drug was removed. The SPN antigen was present in nuclei and mitotic spindles of all human and mammalian cell lines and tissues so far tested. When microinjected into the cytoplasm or nuclei of HeLa cells, one antibody caused a block in mitosis. Total cell number remained constant or decreased slightly after 24 h. At this time, about half the cells were arrested in a prometaphase-like state and revealed aberrant spindles. Many other cells were multinucleate. These results show that the SPN antigen is a protein associated with mitotic spindle microtubules which has to function correctly for the cell to complete mitosis.

MeSH Terms
Antibodies, Monoclonal Antigens, Nuclear Autoantigens/immunology Blotting, Western Fluorescent Antibody Technique HeLa Cells Humans Immunoglobulin G/immunology Microinjections Mitosis Molecular Weight Nocodazole/pharmacology Nuclear Proteins/metabolism Spindle Apparatus/chemistry,drug effects
Chemicals
Antibodies, Monoclonal Antigens, Nuclear Autoantigens Immunoglobulin G Nuclear Proteins Nocodazole
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kallajoki M
Max Planck Institute for Biophysical Chemistry, Department of Biochemistry, Göttingen, FRG.
Weber K
Osborn M
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1991-11-00
Pages
3351-62
Language
English
Region
England
NLM ID
8208664
PMCID
PMC453063
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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