Search tips
Search criteria 


Logo of jcellbiolHomeThe Rockefeller University PressEditorsContactInstructions for AuthorsThis issue
J Cell Biol. 1994 December 1; 127(5): 1301–1310.
PMCID: PMC2120267

Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle


To test the popular but unproven assumption that the metaphase-anaphase transition in vertebrate somatic cells is subject to a checkpoint that monitors chromosome (i.e., kinetochore) attachment to the spindle, we filmed mitosis in 126 PtK1 cells. We found that the time from nuclear envelope breakdown to anaphase onset is linearly related (r2 = 0.85) to the duration the cell has unattached kinetochores, and that even a single unattached kinetochore delays anaphase onset. We also found that anaphase is initiated at a relatively constant 23-min average interval after the last kinetochore attaches, regardless of how long the cell possessed unattached kinetochores. From these results we conclude that vertebrate somatic cells possess a metaphase-anaphase checkpoint control that monitors sister kinetochore attachment to the spindle. We also found that some cells treated with 0.3-0.75 nM Taxol, after the last kinetochore attached to the spindle, entered anaphase and completed normal poleward chromosome motion (anaphase A) up to 3 h after the treatment--well beyond the 9-48-min range exhibited by untreated cells. The fact that spindle bipolarity and the metaphase alignment of kinetochores are maintained in these cells, and that the chromosomes move poleward during anaphase, suggests that the checkpoint monitors more than just the attachment of microtubules at sister kinetochores or the metaphase alignment of chromosomes. Our data are most consistent with the hypothesis that the checkpoint monitors an increase in tension between kinetochores and their associated microtubules as biorientation occurs.

Full Text

The Full Text of this article is available as a PDF (2.5M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Ault JG, Rieder CL. Chromosome mal-orientation and reorientation during mitosis. Cell Motil Cytoskeleton. 1992;22(3):155–159. [PubMed]
  • Bernat RL, Borisy GG, Rothfield NF, Earnshaw WC. Injection of anticentromere antibodies in interphase disrupts events required for chromosome movement at mitosis. J Cell Biol. 1990 Oct;111(4):1519–1533. [PMC free article] [PubMed]
  • Earnshaw WC, Bernat RL, Cooke CA, Rothfield NF. Role of the centromere/kinetochore in cell cycle control. Cold Spring Harb Symp Quant Biol. 1991;56:675–685. [PubMed]
  • Gorbsky GJ, Ricketts WA. Differential expression of a phosphoepitope at the kinetochores of moving chromosomes. J Cell Biol. 1993 Sep;122(6):1311–1321. [PMC free article] [PubMed]
  • Hartwell LH, Weinert TA. Checkpoints: controls that ensure the order of cell cycle events. Science. 1989 Nov 3;246(4930):629–634. [PubMed]
  • Hoyt MA, Totis L, Roberts BT. S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell. 1991 Aug 9;66(3):507–517. [PubMed]
  • Hunt T, Luca FC, Ruderman JV. The requirements for protein synthesis and degradation, and the control of destruction of cyclins A and B in the meiotic and mitotic cell cycles of the clam embryo. J Cell Biol. 1992 Feb;116(3):707–724. [PMC free article] [PubMed]
  • Jensen CG, Davison EA, Bowser SS, Rieder CL. Primary cilia cycle in PtK1 cells: effects of colcemid and taxol on cilia formation and resorption. Cell Motil Cytoskeleton. 1987;7(3):187–197. [PubMed]
  • Jordan MA, Thrower D, Wilson L. Mechanism of inhibition of cell proliferation by Vinca alkaloids. Cancer Res. 1991 Apr 15;51(8):2212–2222. [PubMed]
  • Jordan MA, Thrower D, Wilson L. Effects of vinblastine, podophyllotoxin and nocodazole on mitotic spindles. Implications for the role of microtubule dynamics in mitosis. J Cell Sci. 1992 Jul;102(Pt 3):401–416. [PubMed]
  • Jordan MA, Toso RJ, Thrower D, Wilson L. Mechanism of mitotic block and inhibition of cell proliferation by taxol at low concentrations. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9552–9556. [PubMed]
  • Kung AL, Sherwood SW, Schimke RT. Cell line-specific differences in the control of cell cycle progression in the absence of mitosis. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9553–9557. [PubMed]
  • Li R, Murray AW. Feedback control of mitosis in budding yeast. Cell. 1991 Aug 9;66(3):519–531. [PubMed]
  • Murray AW. Creative blocks: cell-cycle checkpoints and feedback controls. Nature. 1992 Oct 15;359(6396):599–604. [PubMed]
  • Nicklas RB. The forces that move chromosomes in mitosis. Annu Rev Biophys Biophys Chem. 1988;17:431–449. [PubMed]
  • Nicklas RB. Chance encounters and precision in mitosis. J Cell Sci. 1988 Mar;89(Pt 3):283–285. [PubMed]
  • Rattner JB, Berns MW. Centriole behavior in early mitosis of rat kangaroo cells (PTK2). Chromosoma. 1976 Mar 10;54(4):387–395. [PubMed]
  • Rieder CL. Effect of hypothermia (20-25 degrees C) on mitosis in PtK1 cells. Cell Biol Int Rep. 1981 Jun;5(6):563–573. [PubMed]
  • Rieder CL. Formation of the astral mitotic spindle: ultrastructural basis for the centrosome-kinetochore interaction. Electron Microsc Rev. 1990;3(2):269–300. [PubMed]
  • Rieder CL. Mitosis: towards a molecular understanding of chromosome behavior. Curr Opin Cell Biol. 1991 Feb;3(1):59–66. [PubMed]
  • Rieder CL, Alexander SP. The attachment of chromosomes to the mitotic spindle and the production of aneuploidy in newt lung cells. Prog Clin Biol Res. 1989;318:185–194. [PubMed]
  • Rieder CL, Palazzo RE. Colcemid and the mitotic cycle. J Cell Sci. 1992 Jul;102(Pt 3):387–392. [PubMed]
  • Rieder CL, Salmon ED. Motile kinetochores and polar ejection forces dictate chromosome position on the vertebrate mitotic spindle. J Cell Biol. 1994 Feb;124(3):223–233. [PMC free article] [PubMed]
  • Roos UP. Light and electron microscopy of rat kangaroo cells in mitosis. III. Patterns of chromosome behavior during prometaphase. Chromosoma. 1976 Mar 10;54(4):363–385. [PubMed]
  • ROSE G. A separable and multipurpose tissue culture chamber. Tex Rep Biol Med. 1954;12(4):1074–1083. [PubMed]
  • Skibbens RV, Skeen VP, Salmon ED. Directional instability of kinetochore motility during chromosome congression and segregation in mitotic newt lung cells: a push-pull mechanism. J Cell Biol. 1993 Aug;122(4):859–875. [PMC free article] [PubMed]
  • Sluder G. Role of spindle microtubules in the control of cell cycle timing. J Cell Biol. 1979 Mar;80(3):674–691. [PMC free article] [PubMed]
  • Sluder G, Begg DA. Control mechanisms of the cell cycle: role of the spatial arrangement of spindle components in the timing of mitotic events. J Cell Biol. 1983 Sep;97(3):877–886. [PMC free article] [PubMed]
  • Sluder G, Miller FJ, Thompson EA, Wolf DE. Feedback control of the metaphase-anaphase transition in sea urchin zygotes: role of maloriented chromosomes. J Cell Biol. 1994 Jul;126(1):189–198. [PMC free article] [PubMed]
  • Snyder JA, Hamilton BT, Mullins JM. Loss of mitotic centrosomal microtubule initiation capacity at the metaphase-anaphase transition. Eur J Cell Biol. 1982 Jun;27(2):191–199. [PubMed]
  • Spencer F, Hieter P. Centromere DNA mutations induce a mitotic delay in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1992 Oct 1;89(19):8908–8912. [PubMed]
  • Tomkiel J, Cooke CA, Saitoh H, Bernat RL, Earnshaw WC. CENP-C is required for maintaining proper kinetochore size and for a timely transition to anaphase. J Cell Biol. 1994 May;125(3):531–545. [PMC free article] [PubMed]
  • Vandré DD, Borisy GG. Anaphase onset and dephosphorylation of mitotic phosphoproteins occur concomitantly. J Cell Sci. 1989 Oct;94(Pt 2):245–258. [PubMed]
  • Wendell KL, Wilson L, Jordan MA. Mitotic block in HeLa cells by vinblastine: ultrastructural changes in kinetochore-microtubule attachment and in centrosomes. J Cell Sci. 1993 Feb;104(Pt 2):261–274. [PubMed]
  • Zirkle RE. Ultraviolet-microbeam irradiation of newt-cell cytoplasm: spindle destruction, false anaphase, and delay of true anaphase. Radiat Res. 1970 Mar;41(3):516–537. [PubMed]

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