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PMID: 2575618 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Monoclonal antibody against the centrosome.

Journal of cell science ·Vol. 93 ( Pt 1) ·1989-05-00 ·Pages 63-9

Rao PN, Zhao JY, Ganju RK, Ashorn CL

Abstract

A monoclonal antibody, MPM-13, raised against mitotic HeLa cell extracts recognized a perinuclear area in interphase cells and spindle poles in mitotic cells of human, mouse, Chinese hamster and sea urchin. On immunoblots MPM-13 recognized a major protein band at 43 X 10(3) Mr and a variable minor band at 56 X 10(3) Mr in both mitotic and interphase HeLa cells. These antigens were detectable in a variety of mammalian cells as well as in the unicellular ciliate Tetrahymena. In cells arrested in mitosis by colcemid and stained with MPM-13 by indirect immunofluorescence, numerous fluorescent speckles were seen throughout the cytoplasm. Reversal of colcemid block in Chinese hamster ovary (CHO) cells by washing resulted in the reappearance of a fluorescent patch at the poles of the re-formed spindles. In HeLa cells arrested in mitosis by the microtubule stabilizing drug taxol, MPM-13 stained a large fluorescent patch encircled by dark metaphase chromosomes. This pattern indicated the failure of centrioles to move to the opposite poles in the presence of taxol. These data indicate that the MPM-13 antigens are associated with the colcemid-sensitive pericentriolar material from which microtubules originate but not with the centrioles themselves. It is also clear that these antigens are highly conserved during evolution.

MeSH Terms
Alkaloids/pharmacology Animals Antibodies, Monoclonal Cell Line Centrioles/analysis,ultrastructure Demecolcine/pharmacology Electrophoresis, Polyacrylamide Gel Fluorescent Antibody Technique HeLa Cells/analysis,cytology,ultrastructure Humans Immunoblotting Interphase Mitosis Molecular Weight Organelles/analysis,drug effects,ultrastructure Paclitaxel Proteins/isolation & purification
Chemicals
Alkaloids Antibodies, Monoclonal Proteins Paclitaxel Demecolcine
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Rao P N
Department of Medical Oncology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Zhao J Y
Ganju R K
Ashorn C L
Article Info
Journal
Journal of cell science
Abbr.
J Cell Sci
ISSN
0021-9533
Published
1989-05-00
Pages
63-9
Language
English
Region
England
NLM ID
0052457
Subset
IM
Grants
NCI NIH HHS · CA27544 · United States
NCI NIH HHS · CA34783 · United States
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