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. 1981 Dec 1;91(3):822–826. doi: 10.1083/jcb.91.3.822

Microtubule-nucleating activity of centrosomes in Chinese hamster ovary cells is independent of the centriole cycle but coupled to the mitotic cycle

PMCID: PMC2112779  PMID: 7328124

Abstract

The nuclear-centrosome complex was isolated from interphase Chinese hamster ovary (CHO) cells, and, with exogenous brain tubulin as a source of subunits, the centrosome, while attached to the nucleus, was demonstrated to nucleate microtubule formation in vitro. We attempted to quantitate the nucleating activity in order to compare the activity of mitotic and interphase centrosomes. However, the proximity of the nucleus hindered these attempts, and efforts to chemically or mechanically remove the centrosome led to diminished nucleating activity. Therefore, the nuclear-centrosome complex was dissociated biologically through use of the cytochalasin B procedure for enucleation of cells. Cytoplasts were prepared that retained the centrosome. Lysis of the cytoplasts released free centrosomes that could nucleate microtubules in vitro. The nucleating activities of interphase and mitotic centrosomes were compared. In addition, through the use of whole-mount electron microscopy, the configuration of the centrioles was analyzed and the number of microtubules nucleated was determined as a function of the centriole cycle. Nucleating activity did not change discernibly throughout interphase but increased approximately fivefold at the transition to mitosis. Thus, we conclude that the nucleating activity of the centrosome is relatively independent of the centriole cycle but coupled to the mitotic cycle.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Borisy G. G., Marcum J. M., Olmsted J. B., Murphy D. B., Johnson K. A. Purification of tubulin and associated high molecular weight proteins from porcine brain and characterization of microtubule assembly in vitro. Ann N Y Acad Sci. 1975 Jun 30;253:107–132. doi: 10.1111/j.1749-6632.1975.tb19196.x. [DOI] [PubMed] [Google Scholar]
  2. Brinkley B. R., Fuller E. M., Highfield D. P. Cytoplasmic microtubules in normal and transformed cells in culture: analysis by tubulin antibody immunofluorescence. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4981–4985. doi: 10.1073/pnas.72.12.4981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Carter S. B. Effects of cytochalasins on mammalian cells. Nature. 1967 Jan 21;213(5073):261–264. doi: 10.1038/213261a0. [DOI] [PubMed] [Google Scholar]
  4. Den H., Malinzak D. A., Keating H. J., Rosenberg A. Influence of Concanavalin A, wheat germ agglutinin, and soybean agglutinin on the fusion of myoblasts in vitro. J Cell Biol. 1975 Dec;67(3):826–834. doi: 10.1083/jcb.67.3.826. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Frankel F. R. Organization and energy-dependent growth of microtubules in cells. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2798–2802. doi: 10.1073/pnas.73.8.2798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Freed J. J., Lebowitz M. M. The association of a class of saltatory movements with microtubules in cultured cells. J Cell Biol. 1970 May;45(2):334–354. doi: 10.1083/jcb.45.2.334. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Goldman R. D., Pollack R., Chang C. M., Bushnell A. Properties of enucleated cells. III. Changes in cytoplasmic architecture of enucleated BHK21 cells following trypsinization and replating. Exp Cell Res. 1975 Jun;93(1):175–183. doi: 10.1016/0014-4827(75)90437-1. [DOI] [PubMed] [Google Scholar]
  8. Gould R. R., Borisy G. G. The pericentriolar material in Chinese hamster ovary cells nucleates microtubule formation. J Cell Biol. 1977 Jun;73(3):601–615. doi: 10.1083/jcb.73.3.601. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kuriyama R., Borisy G. G. Centriole cycle in Chinese hamster ovary cells as determined by whole-mount electron microscopy. J Cell Biol. 1981 Dec;91(3 Pt 1):814–821. doi: 10.1083/jcb.91.3.814. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Osborn M., Weber K. Cytoplasmic microtubules in tissue culture cells appear to grow from an organizing structure towards the plasma membrane. Proc Natl Acad Sci U S A. 1976 Mar;73(3):867–871. doi: 10.1073/pnas.73.3.867. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Prescott D. M., Myerson D., Wallace J. Enucleation of mammalian cells with cytochalasin B. Exp Cell Res. 1972;71(2):480–485. doi: 10.1016/0014-4827(72)90322-9. [DOI] [PubMed] [Google Scholar]
  12. Robbins E., Jentzsch G., Micali A. The centriole cycle in synchronized HeLa cells. J Cell Biol. 1968 Feb;36(2):329–339. doi: 10.1083/jcb.36.2.329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Snyder J. A., McIntosh J. R. Initiation and growth of microtubules from mitotic centers in lysed mammalian cells. J Cell Biol. 1975 Dec;67(3):744–760. doi: 10.1083/jcb.67.3.744. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Spiegelman B. M., Lopata M. A., Kirschner M. W. Multiple sites for the initiation of microtubule assembly in mammalian cells. Cell. 1979 Feb;16(2):239–252. doi: 10.1016/0092-8674(79)90002-3. [DOI] [PubMed] [Google Scholar]
  15. THEG D. E. CYTOPLASMIC MICROTUBULES IN DIFFERENT ANIMAL CELLS. J Cell Biol. 1964 Nov;23:265–275. doi: 10.1083/jcb.23.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Telzer B. R., Rosenbaum J. L. Cell cycle-dependent, in vitro assembly of microtubules onto pericentriolar material of HeLa cells. J Cell Biol. 1979 Jun;81(3):484–497. doi: 10.1083/jcb.81.3.484. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wise G. E., Prescott D. M. Ultrastructure of enucleated mammalian cells in culture. Exp Cell Res. 1973 Sep;81(1):63–72. doi: 10.1016/0014-4827(73)90111-0. [DOI] [PubMed] [Google Scholar]

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