Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1986 Nov 1;103(5):1863–1872. doi: 10.1083/jcb.103.5.1863

A novel mitotic spindle pole component that originates from the cytoplasm during prophase

PMCID: PMC2114391  PMID: 3536956

Abstract

Several unique aspects of mitotic spindle formation have been revealed by investigation of an autoantibody present in the serum of a patient with the CREST (calcinosis, Raynaud's phenomenon, esophageal dysmotility, schlerodacytyly, and telangiectasias) syndrome. This antibody was previously shown to label at the spindle poles of metaphase and anaphase cells and to be absent from interphase cells. We show here that the serum stained discrete cytoplasmic foci in early prophase cells and only later localized to the spindle poles. The cytoplasmic distribution of the antigen was also seen in nocodazole- arrested cells and prophase cells in populations treated with taxol. In normal and taxol-treated cells, the microtubules appeared to emanate from the cytoplasmic foci and polar stain, and in cells released from nocodazole block, microtubules regrew from antigen-containing centers. This characteristic distribution suggests that the antigen is part of a microtubule organizing center. Thus, we propose that a prophase originating polar antigen functions in spindle pole organization as a coalescing microtubule organizing center that is present only during mitosis. Characterization of the serum showed reactions with multiple proteins at 115, 110, 50, 36, 30, and 28 kD. However, affinity-eluted antibody from the 115/110-kD bands was shown to specifically label the spindle pole and cytosolic foci in prophase cells.

Full Text

The Full Text of this article is available as a PDF (3.5 MB).

Selected References

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

  1. Anderson R. G., Floyd A. K. Electrophoretic analysis of basal body (centriole) proteins. Biochemistry. 1980 Nov 25;19(24):5625–5631. doi: 10.1021/bi00565a026. [DOI] [PubMed] [Google Scholar]
  2. Berlin R. D., Oliver J. M., Walter R. J. Surface functions during Mitosis I: phagocytosis, pinocytosis and mobility of surface-bound Con A. Cell. 1978 Oct;15(2):327–341. doi: 10.1016/0092-8674(78)90002-8. [DOI] [PubMed] [Google Scholar]
  3. Calarco-Gillam P. D., Siebert M. C., Hubble R., Mitchison T., Kirschner M. Centrosome development in early mouse embryos as defined by an autoantibody against pericentriolar material. Cell. 1983 Dec;35(3 Pt 2):621–629. doi: 10.1016/0092-8674(83)90094-6. [DOI] [PubMed] [Google Scholar]
  4. Clayton L., Black C. M., Lloyd C. W. Microtubule nucleating sites in higher plant cells identified by an auto-antibody against pericentriolar material. J Cell Biol. 1985 Jul;101(1):319–324. doi: 10.1083/jcb.101.1.319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cox J. V., Schenk E. A., Olmsted J. B. Human anticentromere antibodies: distribution, characterization of antigens, and effect on microtubule organization. Cell. 1983 Nov;35(1):331–339. doi: 10.1016/0092-8674(83)90236-2. [DOI] [PubMed] [Google Scholar]
  6. Davis F. M., Tsao T. Y., Fowler S. K., Rao P. N. Monoclonal antibodies to mitotic cells. Proc Natl Acad Sci U S A. 1983 May;80(10):2926–2930. doi: 10.1073/pnas.80.10.2926. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Earnshaw W. C., Rothfield N. Identification of a family of human centromere proteins using autoimmune sera from patients with scleroderma. Chromosoma. 1985;91(3-4):313–321. doi: 10.1007/BF00328227. [DOI] [PubMed] [Google Scholar]
  8. Izant J. G., Weatherbee J. A., McIntosh J. R. A microtubule-associated protein in the mitotic spindle and the interphase nucleus. Nature. 1982 Jan 21;295(5846):248–250. doi: 10.1038/295248a0. [DOI] [PubMed] [Google Scholar]
  9. Keryer G., Ris H., Borisy G. G. Centriole distribution during tripolar mitosis in Chinese hamster ovary cells. J Cell Biol. 1984 Jun;98(6):2222–2229. doi: 10.1083/jcb.98.6.2222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Kuriyama R., Keryer G., Borisy G. G. The mitotic spindle of Chinese hamster ovary cells isolated in taxol-containing medium. J Cell Sci. 1984 Mar;66:265–275. doi: 10.1242/jcs.66.1.265. [DOI] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lin W., Fung B., Shyamala M., Kasamatsu H. Identification of antigenically related polypeptides at centrioles and basal bodies. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2373–2377. doi: 10.1073/pnas.78.4.2373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Lydersen B. K., Pettijohn D. E. Human-specific nuclear protein that associates with the polar region of the mitotic apparatus: distribution in a human/hamster hybrid cell. Cell. 1980 Nov;22(2 Pt 2):489–499. doi: 10.1016/0092-8674(80)90359-1. [DOI] [PubMed] [Google Scholar]
  15. Maro B., Howlett S. K., Webb M. Non-spindle microtubule organizing centers in metaphase II-arrested mouse oocytes. J Cell Biol. 1985 Nov;101(5 Pt 1):1665–1672. doi: 10.1083/jcb.101.5.1665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McCarty G. A., Valencia D. W., Fritzler M. J., Barada F. A. A unique antinuclear antibody staining only the mitotic-spindle apparatus. N Engl J Med. 1981 Sep 17;305(12):703–703. doi: 10.1056/NEJM198109173051223. [DOI] [PubMed] [Google Scholar]
  17. Pettijohn D. E., Henzl M., Price C. Nuclear proteins that become part of the mitotic apparatus: a role in nuclear assembly? J Cell Sci Suppl. 1984;1:187–201. doi: 10.1242/jcs.1984.supplement_1.12. [DOI] [PubMed] [Google Scholar]
  18. Price C. M., McCarty G. A., Pettijohn D. E. NuMA protein is a human autoantigen. Arthritis Rheum. 1984 Jul;27(7):774–779. doi: 10.1002/art.1780270708. [DOI] [PubMed] [Google Scholar]
  19. ROBBINS E., GONATAS N. K. THE ULTRASTRUCTURE OF A MAMMALIAN CELL DURING THE MITOTIC CYCLE. J Cell Biol. 1964 Jun;21:429–463. doi: 10.1083/jcb.21.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Roos U. P. Light and electron microscopy of rat kangaroo cells in mitosis. I. Formation and breakdown of the mitotic apparatus. Chromosoma. 1973;40(1):43–82. doi: 10.1007/BF00319836. [DOI] [PubMed] [Google Scholar]
  22. Sager P. R., Brown P. A., Berlin R. D. Analysis of transferrin recycling in mitotic and interphase HeLa cells by quantitative fluorescence microscopy. Cell. 1984 Dec;39(2 Pt 1):275–282. doi: 10.1016/0092-8674(84)90005-9. [DOI] [PubMed] [Google Scholar]
  23. Senécal J. L., Oliver J. M., Rothfield N. Anticytoskeletal autoantibodies in the connective tissue diseases. Arthritis Rheum. 1985 Aug;28(8):889–898. doi: 10.1002/art.1780280808. [DOI] [PubMed] [Google Scholar]
  24. Shyamala M., Atcheson C. L., Kasamatsu H. Stimulation of host centriolar antigen in TC7 cells by simian virus 40: requirement for RNA and protein syntheses and an intact simian virus 40 small-t gene function. J Virol. 1982 Aug;43(2):721–729. doi: 10.1128/jvi.43.2.721-729.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tassin A. M., Maro B., Bornens M. Fate of microtubule-organizing centers during myogenesis in vitro. J Cell Biol. 1985 Jan;100(1):35–46. doi: 10.1083/jcb.100.1.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Turksen K., Aubin J. E., Kalnins V. I. Identification of a centriole-associated protein by antibodies present in normal rabbit sera. Nature. 1982 Aug 19;298(5876):763–765. doi: 10.1038/298763a0. [DOI] [PubMed] [Google Scholar]
  28. Vorobjev I. A., Chentsov YuS Centrioles in the cell cycle. I. Epithelial cells. J Cell Biol. 1982 Jun;93(3):938–949. doi: 10.1083/jcb.93.3.938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Zieve G., Solomon F. Proteins specifically associated with the microtubules of the mammalian mitotic spindle. Cell. 1982 Feb;28(2):233–242. doi: 10.1016/0092-8674(82)90341-5. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES