Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1988 Aug 1;107(2):635–641. doi: 10.1083/jcb.107.2.635

The centrin-based cytoskeleton of Chlamydomonas reinhardtii: distribution in interphase and mitotic cells

PMCID: PMC2115233  PMID: 3047144

Abstract

Monoclonal and polyclonal antibodies raised against algal centrin, a protein of algal striated flagellar roots, were used to characterize the occurrence and distribution of this protein in interphase and mitotic Chlamydomonas cells. Chlamydomonas centrin, as identified by Western immunoblot procedures, is a low molecular (20,000-Mr) acidic protein. Immunofluorescence and immunogold labeling demonstrates that centrin is a component of the distal fiber. In addition, centrin-based flagellar roots link the flagellar apparatus to the nucleus. Two major descending fibers extend from the basal bodies toward the nucleus; each descending fiber branches several times giving rise to 8-16 fimbria which surround and embrace the nucleus. Immunogold labeling indicates that these fimbria are juxtaposed to the outer nuclear envelope. Earlier studies have demonstrated that the centrin-based linkage between the flagellar apparatus and the nucleus is contractile, both in vitro and in living Chlamydomonas cells (Wright, R. L., J. Salisbury, and J. Jarvik. 1985. J. Cell Biol. 101:1903-1912; Salisbury, J. L., M. A. Sanders, and L. Harpst. 1987. J. Cell Biol. 105:1799-1805). Immunofluorescence studies show dramatic changes in distribution of the centrin-based system during mitosis that include a transient contraction at preprophase; division, separation, and re-extension during prophase; and a second transient contraction at the metaphase/anaphase boundary. These observations suggest a fundamental role for centrin in motile events during mitosis.

Full Text

The Full Text of this article is available as a PDF (2.1 MB).

Selected References

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

  1. Ares M., Jr, Howell S. H. Cell cycle stage-specific accumulation of mRNAs encoding tubulin and other polypeptides in Chlamydomonas. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5577–5581. doi: 10.1073/pnas.79.18.5577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cavalier-Smith T. Basal body and flagellar development during the vegetative cell cycle and the sexual cycle of Chlamydomonas reinhardii. J Cell Sci. 1974 Dec;16(3):529–556. doi: 10.1242/jcs.16.3.529. [DOI] [PubMed] [Google Scholar]
  3. Coss R. A. Mitosis in Chlamydomonas reinhardtii basal bodies and the mitotic apparatus. J Cell Biol. 1974 Oct;63(1):325–329. doi: 10.1083/jcb.63.1.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Detmers P. A., Carboni J. M., Condeelis J. Localization of actin in Chlamydomonas using antiactin and NBD-phallacidin. Cell Motil. 1985;5(5):415–430. doi: 10.1002/cm.970050505. [DOI] [PubMed] [Google Scholar]
  5. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay (ELISA). Quantitative assay of immunoglobulin G. Immunochemistry. 1971 Sep;8(9):871–874. doi: 10.1016/0019-2791(71)90454-x. [DOI] [PubMed] [Google Scholar]
  6. Goldsby R. A., Osborne B. A., Simpson E., Herzenberg L. A. Hybrid cell lines with T-cell characteristics. Nature. 1977 Jun 23;267(5613):707–708. doi: 10.1038/267707a0. [DOI] [PubMed] [Google Scholar]
  7. Goldsby R. A., Zipser E. The isolation and replica plating of mammalian cell clones. Exp Cell Res. 1969 Feb;54(2):271–275. doi: 10.1016/0014-4827(69)90250-x. [DOI] [PubMed] [Google Scholar]
  8. Goodenough U. W., Weiss R. L. Interrelationships between microtubules, a striated fiber, and the gametic mating structure of Chlamydomonas reinhardi. J Cell Biol. 1978 Feb;76(2):430–438. doi: 10.1083/jcb.76.2.430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hoops H. J., Witman G. B. Outer doublet heterogeneity reveals structural polarity related to beat direction in Chlamydomonas flagella. J Cell Biol. 1983 Sep;97(3):902–908. doi: 10.1083/jcb.97.3.902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hyams J. S., Borisy G. G. Flagellar coordination in Chlamydomonas reinhardtii: isolation and reactivation of the flagellar apparatus. Science. 1975 Sep 12;189(4206):891–893. doi: 10.1126/science.1098148. [DOI] [PubMed] [Google Scholar]
  11. Hyams J. S., Borisy G. G. Isolated flagellar apparatus of Chlamydomonas: characterization of forward swimming and alteration of waveform and reversal of motion by calcium ions in vitro. J Cell Sci. 1978 Oct;33:235–253. doi: 10.1242/jcs.33.1.235. [DOI] [PubMed] [Google Scholar]
  12. Johnson U. G., Porter K. R. Fine structure of cell division in Chlamydomonas reinhardi. Basal bodies and microtubules. J Cell Biol. 1968 Aug;38(2):403–425. doi: 10.1083/jcb.38.2.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kuchka M. R., Jarvik J. W. Analysis of flagellar size control using a mutant of Chlamydomonas reinhardtii with a variable number of flagella. J Cell Biol. 1982 Jan;92(1):170–175. doi: 10.1083/jcb.92.1.170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. 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]
  15. Mazia D. Centrosomes and mitotic poles. Exp Cell Res. 1984 Jul;153(1):1–15. doi: 10.1016/0014-4827(84)90442-7. [DOI] [PubMed] [Google Scholar]
  16. McFadden G. I., Schulze D., Surek B., Salisbury J. L., Melkonian M. Basal body reorientation mediated by a Ca2+-modulated contractile protein. J Cell Biol. 1987 Aug;105(2):903–912. doi: 10.1083/jcb.105.2.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  18. Pickett-Heaps J. D., Tippit D. H., Porter K. R. Rethinking mitosis. Cell. 1982 Jul;29(3):729–744. doi: 10.1016/0092-8674(82)90435-4. [DOI] [PubMed] [Google Scholar]
  19. Piperno G., Luck D. J. Microtubular proteins of Chlamydomonas reinhardtii. An immunochemical study based on the use of an antibody specific for the beta-tubulin subunit. J Biol Chem. 1977 Jan 10;252(1):383–391. [PubMed] [Google Scholar]
  20. Ringo D. L. Flagellar motion and fine structure of the flagellar apparatus in Chlamydomonas. J Cell Biol. 1967 Jun;33(3):543–571. doi: 10.1083/jcb.33.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. SAGER R., GRANICK S. Nutritional control of sexuality in Chlamydomonas reinhardi. J Gen Physiol. 1954 Jul 20;37(6):729–742. doi: 10.1085/jgp.37.6.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Salisbury J. L., Aebig K. W., Coling D. E. Isolation of the calcium-modulated contractile protein of striated flagellar roots. Methods Enzymol. 1986;134:408–414. doi: 10.1016/0076-6879(86)34107-7. [DOI] [PubMed] [Google Scholar]
  23. Salisbury J. L., Baron A. T., Coling D. E., Martindale V. E., Sanders M. A. Calcium-modulated contractile proteins associated with the eucaryotic centrosome. Cell Motil Cytoskeleton. 1986;6(2):193–197. doi: 10.1002/cm.970060218. [DOI] [PubMed] [Google Scholar]
  24. Salisbury J. L., Baron A., Surek B., Melkonian M. Striated flagellar roots: isolation and partial characterization of a calcium-modulated contractile organelle. J Cell Biol. 1984 Sep;99(3):962–970. doi: 10.1083/jcb.99.3.962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Salisbury J. L., Floyd G. L. Calcium-induced contraction of the rhizoplast of a quadriflagellate green alga. Science. 1978 Dec 1;202(4371):975–977. doi: 10.1126/science.202.4371.975. [DOI] [PubMed] [Google Scholar]
  26. Salisbury J. L., Sanders M. A., Harpst L. Flagellar root contraction and nuclear movement during flagellar regeneration in Chlamydomonas reinhardtii. J Cell Biol. 1987 Oct;105(4):1799–1805. doi: 10.1083/jcb.105.4.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Wright R. L., Salisbury J., Jarvik J. W. A nucleus-basal body connector in Chlamydomonas reinhardtii that may function in basal body localization or segregation. J Cell Biol. 1985 Nov;101(5 Pt 1):1903–1912. doi: 10.1083/jcb.101.5.1903. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES