Skip to main content
Springer Nature Link
Log in

Relevance of kinetochore size and microtubule-binding capacity for stable chromosome attachment during mitosis in PtK1 cells

  • Published:
Chromosome Research Aims and scope Submit manuscript

Abstract

Chromosomes attach to the mitotic spindle via their kinetochores. The average number of spindle microtubules binding to each kinetochore varies with species, the stage of mitosis, and the length of time that the kinetochore has been attached to the spindle. In this report, we investigate how kinetochore microtubule number varies with kinetochore size and chromosome size in PtK1 cells. From an analysis of serial-section electron micrographs, we determined that the average surface area of metaphase, taxol-treated metaphase, and anaphase kinetochores is 0.16 ± 0.05 μm2 (N = 181). Surprisingly, kinetochore microtubules are packed more densely on the smaller kinetochores, as seen by a reduction in the average spacing between kinetochore microtubules from 89 nm to 59 nm. Our interpretation of this result is that PtK1 cells require a minimum kinetochore microtubule-binding capacity for survival during repeated rounds of mitotic division. We estimate the lower limit to be 23 kinetochore microtubules and suggest that this capacity is required to ensure stable attachment during the dynamic and highly stochastic process of kinetochore fiber formation. There is a modest but statistically significant increase in kinetochore microtubule number with chromosome size, indicating that chromosome size is a minor determinant of kinetochore microtubule number.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Belmont LD, Hyman AA, Sawin KE, Mitchison TJ (1990) Realtime visualization of cell cycle-dependent changes in microtubule dynamics in cytoplasmic extracts. Cell 62: 579–589.

    Google Scholar 

  • Bickel PJ, Doksum KA (1977) Mathematical Statistics: Basic Ideas & Selected Topics. Oakland, CA: Holden Day.

    Google Scholar 

  • Carminati JL, Stearns T (1997) Microtubules orient the mitotic spindle in yeast through dynein-dependent interactions with the cell cortex. J Cell Biol 138: 629–641.

    Google Scholar 

  • Cassimeris L, Rieder CL, Rupp G, Salmon ED (1990) Stability of microtubule attachment to metaphase kinetochores in PtK1 cells. J Cell Sci 96: 9–15.

    Google Scholar 

  • Cherry LM, Johnston DA (1987) Size variation in kinetochores of human chromosomes. Hum Genet 75: 155–158.

    Google Scholar 

  • Cherry LM, Faulkner AJ, Grossberg LA, Balczon R (1989) Kinetochore size variation in mammalian chromosomes: an image analysis study with evolutionary implications. J Cell Sci 92: 281–289.

    Google Scholar 

  • Ding R, McDonald KL, McIntosh R (1993) Three-dimensional reconstruction and analysis of mitotic spindles from the yeast, Schizosaccharomyces pombe. J Cell Biol 120: 141–151.

    Google Scholar 

  • Frank J, Radermacher M, Penczek P et al. (1996) SPIDER and WEB: processing and visualization of images in 3D electron microscopy and related fields. J Struct Biol 116: 190–199.

    Google Scholar 

  • Fuge H (1978) Ultrastructure of the mitotic spindle. Int Rev Cytol 6(Suppl.): 1–58.

    Google Scholar 

  • Gorbsky GJ, Sammak PJ, Borisy GG (1988) Microtubule dynamics and chromosome motion visualized in living anaphase cells. J Cell Biol 106: 1185–1192.

    Google Scholar 

  • Hayden JH, Bowser, SS, Rieder, CL (1990) Kinetochores capture astral microtubules during chromosome attachment to the mitotic spindle: direct visualization in live newt lung cells. J Cell Biol 111: 1039–1045.

    Google Scholar 

  • Hays TS, Salmon ED (1990) Poleward force at the kinetochore in metaphase depends on the number of kinetochore microtubules. J Cell Biol 110: 391–404.

    Google Scholar 

  • Inoué S, Salmon ED (1995) Force generation by microtubule assembly/disassembly in mitosis and related movements. Mol Biol Cell 6: 1619–1640.

    Google Scholar 

  • Jensen CG (1982) Dynamics of spindle microtubule organization: kinetochore fiber microtubules of plant endosperm. J Cell Biol 92: 540–558.

    Google Scholar 

  • Levan A, Nichols WW, Peluse M, Coriell LL (1966) The stemline chromosomes of three cell lines representing different vertebrate classes. Chromosoma (Berl) 18: 343–358.

    Google Scholar 

  • Levan G (1970) Contributions to the chromosomal characterization of the PTK 1 rat-kangaroo cell line. Hereditas 64: 85–96.

    Google Scholar 

  • Li X, Nicklas RB (1995) Mitotic forces control a cell-cycle checkpoint. Nature 373: 630–632.

    Google Scholar 

  • Lin H-P P, Ault JG, Church K (1981) Meiosis in Drosophila melanogaster. I. Chromosome identification and kinetochore microtubule numbers during the first and second meiotic divisions in males. Chromosoma (Berl) 83: 507–521.

    Google Scholar 

  • McDonald KL, O'Toole ET, Mastronarde DN, McIntosh JR (1992) Kinetochore microtubules in PTK cells. J Cell Biol 118: 369–383.

    Google Scholar 

  • McEwen BF, Heagle AB, Cassels GO, Buttle KF, Rieder CL (1997) Kinetochore fiber maturation in PtK1 cells and its implications for the mechanisms of chromosome congression and anaphase onset. J Cell Biol 137: 1567–1580.

    Google Scholar 

  • Mitchison TJ, Salmon ED (1992) Poleward kinetochore fiber movement occurs during both metaphase and anaphase-A in newt lung cell mitosis. J Cell Biol 119: 569–582.

    Google Scholar 

  • Moens PB (1978) Kinetochores of grasshoppers with Robertsonian chromosome fusions. Chromosoma (Berl) 67: 41–54.

    Google Scholar 

  • Moens PB (1979) Kinetochore microtubule numbers of different sized chromosomes. J Cell Biol 83: 556–561.

    Google Scholar 

  • Nicklas RB (1989) The motor for poleward chromosome movement in anaphase is in or near the kinetochore. J Cell Biol 109: 2245–2255.

    Google Scholar 

  • Pearson ES, Hartley HO (1972) Biometrika Tables for Statisticians, Vol. I, 3rdedn. Cambridge: Cambridge University Press.

    Google Scholar 

  • Peterson JB, Ris H (1976) Electron microscopic study of the spindle and chromosome movement in the yeast Saccharomyces cerevisiae. J Cell Sci 22: 219–242.

    Google Scholar 

  • Pierre RV (1985) Age and loss of the Y chromosome. In: Sanberg AA, ed. The Y Chromosome, pt B: Clinical aspects of Y Chromosome Abnormalities. New York: Liss, pp 53–60.

    Google Scholar 

  • Rieder CL (1982) The formation, structure and composition of the mammalian kinetochore and kinetochore fiber. Int Rev Cytol 79: 1–58.

    Google Scholar 

  • Rieder CL, Alexander SP (1990) Kinetochores are transported poleward along a single astral microtubule during chromosome attachment to the spindle in newt lung cells. J Cell Biol 110: 81–95.

    Google Scholar 

  • Rieder CL, Salmon ED (1994) Motile kinetochores and polar ejection forces dictate chromosome position on the vertebrate mitotic spindle. J Cell Biol 124: 223–233.

    Google Scholar 

  • Rieder CL, Schultz A, Cole R, Sluder G (1994) Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle. J Cell Biol 127: 1301–1310.

    Google Scholar 

  • Rieder CL, Cole RW, Khodjakov A, Sluder G (1995) The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores. J Cell Biol 130: 941–948.

    Google Scholar 

  • Roos U-P (1973) Light and electron microscopy of rat kangaroo cells in mitosis. II. Kinetochore structure and function. Chromosoma (Berl) 41: 195–220.

    Google Scholar 

  • Skibbens RV, Petrie Skeen V, Salmon ED (1993) Directional instability of kinetochore motility during chromosome congression and segregation in mitotic newt lung cells: a push-pull mechanism. J Cell Biol 122: 859–875.

    Google Scholar 

  • Wendell KL, Wilson L, Jordan MA (1993) Mitotic block in HeLa cells by vinblastine: ultrastructural changes in kinetochore-microtubule attachment and in centrosomes. J Cell Sci 104: 261–274.

    Google Scholar 

  • Zhai Y, Kronebusch PJ, Borisy GG (1995) Kinetochore microtubule dynamics and the metaphase-anaphase transition. J Cell Biol 131: 721–734.

    Google Scholar 

  • Zhang D, Nicklas RB (1997) Chromosome congression in mini-spindles containing a single chromosome. Mol Biol Cell 8: 171a.

    Google Scholar 

  • Zinkowski RP, Meyne J, Brinkley BR (1991) The centromere-kinetochore complex: a repeat subunit model. J Cell Biol 113: 1091–1110.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Molecular Medicine, Wadsworth Center, New York State Department of Health, PO Box 509, Albany, NY, 12201-0509 New York, USA

    Bruce F. McEwen, Ye Ding & Amy B. Heagle

Authors
  1. Bruce F. McEwen
    View author publications

    You can also search for this author inPubMed Google Scholar

  2. Ye Ding
    View author publications

    You can also search for this author inPubMed Google Scholar

  3. Amy B. Heagle
    View author publications

    You can also search for this author inPubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

McEwen, B.F., Ding, Y. & Heagle, A.B. Relevance of kinetochore size and microtubule-binding capacity for stable chromosome attachment during mitosis in PtK1 cells. Chromosome Res 6, 123–132 (1998). https://doi.org/10.1023/A:1009239013215

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1009239013215