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. 1987 May;84(9):2771–2775. doi: 10.1073/pnas.84.9.2771

Kinesin is associated with a nonmicrotubule component of sea urchin mitotic spindles.

R J Leslie, R B Hird, L Wilson, J R McIntosh, J M Scholey
PMCID: PMC304740  PMID: 3106977

Abstract

Sea urchin embryos in second division have been lysed into microtubule-stabilizing buffers to yield mitotic cytoskeletons (MCSs) that consist of two mitotic spindles surrounded by a cortical array of filaments. Microtubules have been completely extracted from MCSs by incubation at 0 degrees C with Ca2+-containing buffer. An antibody to the microtubule translocator kinesin stains the spindles in MCSs and in MCSs treated with 5 mM ATP and also stains spindle-remnants of the MCSs after the microtubules have been extracted. We conclude that kinesin binds to a nonmicrotubule component in the mitotic spindle. Based on these results, we present several models of kinesin function in the spindle.

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Selected References

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

  1. Bajer A., Allen R. D. Role of phragmoplast filaments in cell-plate formation. J Cell Sci. 1966 Dec;1(4):455–462. doi: 10.1242/jcs.1.4.455. [DOI] [PubMed] [Google Scholar]
  2. Brady S. T., Lasek R. J., Allen R. D. Fast axonal transport in extruded axoplasm from squid giant axon. Science. 1982 Dec 10;218(4577):1129–1131. doi: 10.1126/science.6183745. [DOI] [PubMed] [Google Scholar]
  3. Branton D., Cohen C. M., Tyler J. Interaction of cytoskeletal proteins on the human erythrocyte membrane. Cell. 1981 Apr;24(1):24–32. doi: 10.1016/0092-8674(81)90497-9. [DOI] [PubMed] [Google Scholar]
  4. Cande W. Z., McDonald K. L. In vitro reactivation of anaphase spindle elongation using isolated diatom spindles. Nature. 1985 Jul 11;316(6024):168–170. doi: 10.1038/316168a0. [DOI] [PubMed] [Google Scholar]
  5. Euteneuer U., McIntosh J. R. Structural polarity of kinetochore microtubules in PtK1 cells. J Cell Biol. 1981 May;89(2):338–345. doi: 10.1083/jcb.89.2.338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fuge H. Ultrastructure of the mitotic spindle. Int Rev Cytol Suppl. 1977;(6):1–58. [PubMed] [Google Scholar]
  7. Harris P. The role of membranes in the ogranization of the mitotic apparatus. Exp Cell Res. 1975 Sep;94(2):409–425. doi: 10.1016/0014-4827(75)90507-8. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Leslie R. J., Pickett-Heaps J. D. Ultraviolet microbeam irradiations of mitotic diatoms: investigation of spindle elongation. J Cell Biol. 1983 Feb;96(2):548–561. doi: 10.1083/jcb.96.2.548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. McDonald K. L., Edwards M. K., McIntosh J. R. Cross-sectional structure of the central mitotic spindle of Diatoma vulgare. Evidence for specific interactions between antiparallel microtubules. J Cell Biol. 1979 Nov;83(2 Pt 1):443–461. doi: 10.1083/jcb.83.2.443. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mitchison T. J., Kirschner M. W. Properties of the kinetochore in vitro. II. Microtubule capture and ATP-dependent translocation. J Cell Biol. 1985 Sep;101(3):766–777. doi: 10.1083/jcb.101.3.766. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. O'Farrell P. Z., Goodman H. M., O'Farrell P. H. High resolution two-dimensional electrophoresis of basic as well as acidic proteins. Cell. 1977 Dec;12(4):1133–1141. doi: 10.1016/0092-8674(77)90176-3. [DOI] [PubMed] [Google Scholar]
  13. OSTERGREN G., MOLE-BAJER J., BAJER A. An interpretation of transport phenomena at mitosis. Ann N Y Acad Sci. 1960 Oct 7;90:381–408. doi: 10.1111/j.1749-6632.1960.tb23258.x. [DOI] [PubMed] [Google Scholar]
  14. Olmsted J. B. Affinity purification of antibodies from diazotized paper blots of heterogeneous protein samples. J Biol Chem. 1981 Dec 10;256(23):11955–11957. [PubMed] [Google Scholar]
  15. 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]
  16. Rebhun L. I. Polarized intracellular particle transport: saltatory movements and cytoplasmic streaming. Int Rev Cytol. 1972;32:93–137. doi: 10.1016/s0074-7696(08)60339-3. [DOI] [PubMed] [Google Scholar]
  17. Rebhun L. I. Structural aspects of saltatory particle movement. J Gen Physiol. 1967 Jul;50(6 Suppl):223–239. doi: 10.1085/jgp.50.6.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Salmon E. D. Mitotic spindles isolated from sea urchin eggs with EGTA lysis buffers. Methods Cell Biol. 1982;25(Pt B):69–105. doi: 10.1016/s0091-679x(08)61421-1. [DOI] [PubMed] [Google Scholar]
  19. Salmon E. D., Segall R. R. Calcium-labile mitotic spindles isolated from sea urchin eggs (Lytechinus variegatus). J Cell Biol. 1980 Aug;86(2):355–365. doi: 10.1083/jcb.86.2.355. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Scholey J. M., Porter M. E., Grissom P. M., McIntosh J. R. Identification of kinesin in sea urchin eggs, and evidence for its localization in the mitotic spindle. Nature. 1985 Dec 5;318(6045):483–486. doi: 10.1038/318483a0. [DOI] [PubMed] [Google Scholar]
  21. 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]
  22. Vale R. D., Reese T. S., Sheetz M. P. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell. 1985 Aug;42(1):39–50. doi: 10.1016/s0092-8674(85)80099-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vale R. D., Schnapp B. J., Mitchison T., Steuer E., Reese T. S., Sheetz M. P. Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro. Cell. 1985 Dec;43(3 Pt 2):623–632. doi: 10.1016/0092-8674(85)90234-x. [DOI] [PubMed] [Google Scholar]
  24. Vale R. D., Schnapp B. J., Reese T. S., Sheetz M. P. Organelle, bead, and microtubule translocations promoted by soluble factors from the squid giant axon. Cell. 1985 Mar;40(3):559–569. doi: 10.1016/0092-8674(85)90204-1. [DOI] [PubMed] [Google Scholar]
  25. Williams R. C., Jr, Lee J. C. Preparation of tubulin from brain. Methods Enzymol. 1982;85(Pt B):376–385. doi: 10.1016/0076-6879(82)85038-6. [DOI] [PubMed] [Google Scholar]

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