Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1984 Dec 1;99(6):2108–2113. doi: 10.1083/jcb.99.6.2108

Reformation of the marginal band of avian erythrocytes in vitro using calf-brain tubulin: peripheral determinants of microtubule form

PMCID: PMC2113563  PMID: 6542106

Abstract

The microtubules of nucleated erythrocytes form an extraordinary structure: they are organized into a marginal band at the periphery of the cell. This unusual organelle, recurring in detail in each cell, provides an excellent opportunity to study the determinants of microtubule form. We have been able to reform the marginal band, using detergent-extracted erythrocytes that have been depleted of microtubules in vivo and phosphocellulose-purified tubulin from calf brain. We find that detergent-extracted cytoskeletons incubated under these conditions again have microtubules, and that the pattern of these microtubules recapitulates several features of the intact marginal band. In particular, most of the microtubules after regrowth are located in a band at the periphery of the cell, and curve to form an ellipse. These results support the hypothesis that the specification of microtubule location and shape in these cells is governed by determinants that reside at the periphery of the cell.

Full Text

The Full Text of this article is available as a PDF (2.4 MB).

Selected References

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

  1. Bertolini B., Monaco G. The microtubule marginal band of the newt erythrocyte. Observations on the isolated band. J Ultrastruct Res. 1976 Jan;54(1):59–67. doi: 10.1016/s0022-5320(76)80008-1. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Brinkley B. R., Cox S. M., Pepper D. A., Wible L., Brenner S. L., Pardue R. L. Tubulin assembly sites and the organization of cytoplasmic microtubules in cultured mammalian cells. J Cell Biol. 1981 Sep;90(3):554–562. doi: 10.1083/jcb.90.3.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cohen W. D., Bartelt D., Jaeger R., Langford G., Nemhauser I. The cytoskeletal system of nucleated erythrocytes. I. Composition and function of major elements. J Cell Biol. 1982 Jun;93(3):828–828. doi: 10.1083/jcb.93.3.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen W. D. Observations of the marginal band system of nucleated erythrocytes. J Cell Biol. 1978 Jul;78(1):260–273. doi: 10.1083/jcb.78.1.260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Duerr A., Pallas D., Solomon F. Molecular analysis of cytoplasmic microtubules in situ: identification of both widespread and specific proteins. Cell. 1981 Apr;24(1):203–211. doi: 10.1016/0092-8674(81)90516-x. [DOI] [PubMed] [Google Scholar]
  7. Goniakowska-Witalińska L., Witaliński W. Evidence for a correlation between the number of marginal band microtubules and the size of vertebrate erthrocytes. J Cell Sci. 1976 Nov;22(2):397–401. doi: 10.1242/jcs.22.2.397. [DOI] [PubMed] [Google Scholar]
  8. Granger B. L., Lazarides E. Structural associations of synemin and vimentin filaments in avian erythrocytes revealed by immunoelectron microscopy. Cell. 1982 Aug;30(1):263–275. doi: 10.1016/0092-8674(82)90032-0. [DOI] [PubMed] [Google Scholar]
  9. Hoffman P. N., Griffin J. W., Price D. L. Control of axonal caliber by neurofilament transport. J Cell Biol. 1984 Aug;99(2):705–714. doi: 10.1083/jcb.99.2.705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Irie S., Sezaki M., Kato Y. A faithful double stain of proteins in the polyacrylamide gels with Coomassie blue and silver. Anal Biochem. 1982 Nov 1;126(2):350–354. doi: 10.1016/0003-2697(82)90526-7. [DOI] [PubMed] [Google Scholar]
  11. Izant J. G., Weatherbee J. A., McIntosh J. R. A microtubule-associated protein antigen unique to mitotic spindle microtubules in PtK1 cells. J Cell Biol. 1983 Feb;96(2):424–434. doi: 10.1083/jcb.96.2.424. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Morrissey J. H. Silver stain for proteins in polyacrylamide gels: a modified procedure with enhanced uniform sensitivity. Anal Biochem. 1981 Nov 1;117(2):307–310. doi: 10.1016/0003-2697(81)90783-1. [DOI] [PubMed] [Google Scholar]
  13. Murphy D. B., Wallis K. T. Brain and erythrocyte microtubules from chicken contain different beta-tubulin polypeptides. J Biol Chem. 1983 Jun 25;258(12):7870–7875. [PubMed] [Google Scholar]
  14. Murray J. M. Disassembly and reconstitution of a membrane-microtubule complex. J Cell Biol. 1984 Apr;98(4):1481–1487. doi: 10.1083/jcb.98.4.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Nachmias V. T., Sullender J., Fallon J., Asch A. Observations on the "cytoskeleton" of human platelets. Thromb Haemost. 1980 Feb 29;42(5):1661–1666. [PubMed] [Google Scholar]
  16. Pepper D. A., Brinkley B. R. Microtubule initiation at kinetochores and centrosomes in lysed mitotic cells. Inhibition of site-specific nucleation by tubulin antibody. J Cell Biol. 1979 Aug;82(2):585–591. doi: 10.1083/jcb.82.2.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Schiff P. B., Fant J., Horwitz S. B. Promotion of microtubule assembly in vitro by taxol. Nature. 1979 Feb 22;277(5698):665–667. doi: 10.1038/277665a0. [DOI] [PubMed] [Google Scholar]
  18. Schiff P. B., Horwitz S. B. Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1561–1565. doi: 10.1073/pnas.77.3.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Solomon F., Magendantz M., Salzman A. Identification with cellular microtubules of one of the co-assemlbing microtubule-associated proteins. Cell. 1979 Oct;18(2):431–438. doi: 10.1016/0092-8674(79)90062-x. [DOI] [PubMed] [Google Scholar]
  20. Stephens R. E. Primary structural differences among tubulin subunits from flagella, cilia, and the cytoplasm. Biochemistry. 1978 Jul 11;17(14):2882–2891. doi: 10.1021/bi00607a029. [DOI] [PubMed] [Google Scholar]
  21. Weingarten M. D., Lockwood A. H., Hwo S. Y., Kirschner M. W. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. 1975 May;72(5):1858–1862. doi: 10.1073/pnas.72.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. 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