Assembly of Microtubules Onto Kinetochores of Isolated Mitotic Chromosomes of HeLa Cells

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1975 Oct 1

PMID 1060085

Citations 45

Authors

B R Telzer

M J MOSES

J L Rosenbaum

Affiliations

Soon will be listed here.

Abstract

The kinetochores of isolated HeLa cell chromosomes attached to an electron microscope specimen grid, fixed in formaldehyde, and stained with alcoholic phosphotungstic acid are visible as dark, preferentially stained structures distinct from the chromatin with which they are associated. When unfixed chromosomes are immobilized by attachment to grids and incubated with chick brain tubulin, microtubules are observed to assemble onto the kinetochores. This demonstrates the competence of kinetochores in isolated chromosomes to act in vitro as microtubule assembly sites and suggests that they also possess this capacity in vivo. In addition, the results provide a possible means for isolating and characterizing kinetochores.

Citing Articles

Centromeric chromatin clearings demarcate the site of kinetochore formation.

Kixmoeller K, Tarasovetc E, Mer E, Chang Y, Black B Cell. 2025; 188(5):1280-1296.e19.

PMID: 39855195 PMC: 11890969. DOI: 10.1016/j.cell.2024.12.025.

Centromeric chromatin clearings demarcate the site of kinetochore formation.

Kixmoeller K, Chang Y, Black B bioRxiv. 2024; .

PMID: 38712116 PMC: 11071481. DOI: 10.1101/2024.04.26.591177.

Microtubule nucleation from the fibrous corona by LIC1-pericentrin promotes chromosome congression.

Wu J, Larreategui-Aparicio A, Lambers M, Bodor D, Klaasen S, Tollenaar E Curr Biol. 2023; 33(5):912-925.e6.

PMID: 36720222 PMC: 10017265. DOI: 10.1016/j.cub.2023.01.010.

Self-organization of kinetochore-fibers in human mitotic spindles.

Conway W, Kiewisz R, Fabig G, Kelleher C, Wu H, Anjur-Dietrich M Elife. 2022; 11.

PMID: 35876665 PMC: 9398449. DOI: 10.7554/eLife.75458.

A celebration of the 25th anniversary of chromatin-mediated spindle assembly.

Verma V, Maresca T Mol Biol Cell. 2022; 33(2):rt1.

PMID: 35076260 PMC: 9236140. DOI: 10.1091/mbc.E21-08-0400.

References

Roth L, DANIELS E . Electron microscopic studies of mitosis in amebae. II. The giant ameba Pelomyxa carolinensis. J Cell Biol. 1962; 12:57-78. PMC: 2106018. DOI: 10.1083/jcb.12.1.57. View

DENTLER W, Granett S, Witman G, Rosenbaum J . Directionality of brain microtubule assembly in vitro. Proc Natl Acad Sci U S A. 1974; 71(5):1710-4. PMC: 388308. DOI: 10.1073/pnas.71.5.1710. View

Weisenberg R, Rosenfeld A . In vitro polymerization of microtubules into asters and spindles in homogenates of surf clam eggs. J Cell Biol. 1975; 64(1):146-58. PMC: 2109483. DOI: 10.1083/jcb.64.1.146. View

Weisenberg R . Microtubule formation in vitro in solutions containing low calcium concentrations. Science. 1972; 177(4054):1104-5. DOI: 10.1126/science.177.4054.1104. View

Roos U . Light and electron microscopy of rat kangaroo cells in mitosis. II. Kinetochore structure and function. Chromosoma. 1973; 41(2):195-220. DOI: 10.1007/BF00319696. View

Burkholder G, OKADA T, Comings D . Whole mount electron microscopy of metaphase. I. Chromosomes and microtubules from mouse oocytes. Exp Cell Res. 1972; 75(2):497-511. DOI: 10.1016/0014-4827(72)90458-2. View

Cande W, Snyder J, Smith D, Summers K, McIntosh J . A functional mitotic spindle prepared from mammalian cells in culture. Proc Natl Acad Sci U S A. 1974; 71(4):1559-63. PMC: 388270. DOI: 10.1073/pnas.71.4.1559. View

Allen C, Borisy G . Structural polarity and directional growth of microtubules of Chlamydomonas flagella. J Mol Biol. 1974; 90(2):381-402. DOI: 10.1016/0022-2836(74)90381-7. View

MOSES M, Counce S . Electron microscopy of kinetochores in whole mount spreads of mitotic chromosomes from hela cells. J Exp Zool. 1974; 189(1):115-20. DOI: 10.1002/jez.1401890110. View

10.

Snell W, DENTLER W, Haimo L, Binder L, Rosenbaum J . Assembly of chick brain tubulin onto isolated basal bodies of Chlamydomonas reinhardi. Science. 1974; 185(4148):357-60. DOI: 10.1126/science.185.4148.357. View

11.

REBHUN L, Rosenbaum J, Lefebvre P, Smith G . Reversible restoration of the birefringence of cold-treated, isolated mitotic apparatus of surf clam eggs with chick brain tubulin. Nature. 1974; 249(453):113-5. DOI: 10.1038/249113a0. View

12.

Counce S, Meyer G . Differentiation of the synaptonemal complex and the kinetochore in Locusta spermatocytes studied by whole mount electron microscopy. Chromosoma. 1973; 44(2):231-53. DOI: 10.1007/BF00329119. View

13.

Comings D, OKADA T . Holocentric chromosomes in Oncopeltus: kinetochore plates are present in mitosis but absent in meiosis. Chromosoma. 1972; 37(2):177-92. DOI: 10.1007/BF00284937. View

14.

Jokelainen P . The ultrastructure and spatial organization of the metaphase kinetochore in mitotic rat cells. J Ultrastruct Res. 1967; 19(1):19-44. DOI: 10.1016/s0022-5320(67)80058-3. View

15.

Binder L, DENTLER W, Rosenbaum J . Assembly of chick brain tubulin onto flagellar microtubules from Chlamydomonas and sea urchin sperm. Proc Natl Acad Sci U S A. 1975; 72(3):1122-6. PMC: 432478. DOI: 10.1073/pnas.72.3.1122. View

16.

MOSES M, Counce S, Paulson D . Synaptonemal complex complement of man in spreads of spermatocytes, with details of the sex chromosome pair. Science. 1975; 187(4174):363-5. View

17.

Wray W, STUBBLEFIELD E . A new method for the rapid isolation of chromosomes, mitotic apparatus, or nuclei from mammalian fibroblasts at near neutral pH. Exp Cell Res. 1970; 59(3):469-78. DOI: 10.1016/0014-4827(70)90656-7. View

18.

MILLER Jr O, Beatty B . Visualization of nucleolar genes. Science. 1969; 164(3882):955-7. DOI: 10.1126/science.164.3882.955. View

19.

ABUELO J, Moore D . The human chromosome. Electron microscopic observations on chromatin fiber organization. J Cell Biol. 1969; 41(1):73-90. PMC: 2107724. DOI: 10.1083/jcb.41.1.73. View

20.

BUCK R . Mitosis and meiosis in Rhodnius prolixus: the fine structure of the spindle and diffuse kinetochore. J Ultrastruct Res. 1967; 18(5):489-501. DOI: 10.1016/s0022-5320(67)80199-0. View