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. 1996 Apr 1;133(1):75–84. doi: 10.1083/jcb.133.1.75

Aberrantly segregating centromeres activate the spindle assembly checkpoint in budding yeast

PMCID: PMC2120768  PMID: 8601615

Abstract

The spindle assembly checkpoint is the mechanism or set of mechanisms that prevents cells with defects in chromosome alignment or spindle assembly from passing through mitosis. We have investigated the effects of mini-chromosomes on this checkpoint in budding yeast by performing pedigree analysis. This method allowed us to observe the frequency and duration of cell cycle delays in individual cells. Short, centromeric linear mini-chromosomes, which have a low fidelity of segregation, cause frequent delays in mitosis. Their circular counterparts and longer linear mini-chromosomes, which segregate more efficiently, show a much lower frequency of mitotic delays, but these delays occur much more frequently in divisions where the mini-chromosome segregates to only one of the two daughter cells. Using a conditional centromere to increase the copy number of a circular mini-chromosome greatly increases the frequency of delayed divisions. In all cases the division delays are completely abolished by the mad mutants that inactivate the spindle assembly checkpoint, demonstrating that the Mad gene products are required to detect the subtle defects in chromosome behavior that have been observed to arrest higher eukaryotic cells in mitosis.

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Selected References

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  1. Aparicio O. M., Billington B. L., Gottschling D. E. Modifiers of position effect are shared between telomeric and silent mating-type loci in S. cerevisiae. Cell. 1991 Sep 20;66(6):1279–1287. doi: 10.1016/0092-8674(91)90049-5. [DOI] [PubMed] [Google Scholar]
  2. Bernat R. L., Borisy G. G., Rothfield N. F., Earnshaw W. C. Injection of anticentromere antibodies in interphase disrupts events required for chromosome movement at mitosis. J Cell Biol. 1990 Oct;111(4):1519–1533. doi: 10.1083/jcb.111.4.1519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cassimeris L., Rieder C. L., Salmon E. D. Microtubule assembly and kinetochore directional instability in vertebrate monopolar spindles: implications for the mechanism of chromosome congression. J Cell Sci. 1994 Jan;107(Pt 1):285–297. doi: 10.1242/jcs.107.1.285. [DOI] [PubMed] [Google Scholar]
  4. Chlebowicz-Sledziewska E., Sledziewski A. Z. Construction of multicopy yeast plasmids with regulated centromere function. Gene. 1985;39(1):25–31. doi: 10.1016/0378-1119(85)90103-9. [DOI] [PubMed] [Google Scholar]
  5. Clarke L., Carbon J. Isolation of a yeast centromere and construction of functional small circular chromosomes. Nature. 1980 Oct 9;287(5782):504–509. doi: 10.1038/287504a0. [DOI] [PubMed] [Google Scholar]
  6. Dani G. M., Zakian V. A. Mitotic and meiotic stability of linear plasmids in yeast. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3406–3410. doi: 10.1073/pnas.80.11.3406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Ferguson B. M., Fangman W. L. A position effect on the time of replication origin activation in yeast. Cell. 1992 Jan 24;68(2):333–339. doi: 10.1016/0092-8674(92)90474-q. [DOI] [PubMed] [Google Scholar]
  8. Futcher B., Carbon J. Toxic effects of excess cloned centromeres. Mol Cell Biol. 1986 Jun;6(6):2213–2222. doi: 10.1128/mcb.6.6.2213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Glotzer M., Murray A. W., Kirschner M. W. Cyclin is degraded by the ubiquitin pathway. Nature. 1991 Jan 10;349(6305):132–138. doi: 10.1038/349132a0. [DOI] [PubMed] [Google Scholar]
  10. Guacci V., Hogan E., Koshland D. Chromosome condensation and sister chromatid pairing in budding yeast. J Cell Biol. 1994 May;125(3):517–530. doi: 10.1083/jcb.125.3.517. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hartwell L. H., Kastan M. B. Cell cycle control and cancer. Science. 1994 Dec 16;266(5192):1821–1828. doi: 10.1126/science.7997877. [DOI] [PubMed] [Google Scholar]
  12. Hartwell L. H., Weinert T. A. Checkpoints: controls that ensure the order of cell cycle events. Science. 1989 Nov 3;246(4930):629–634. doi: 10.1126/science.2683079. [DOI] [PubMed] [Google Scholar]
  13. Hershko A., Ganoth D., Pehrson J., Palazzo R. E., Cohen L. H. Methylated ubiquitin inhibits cyclin degradation in clam embryo extracts. J Biol Chem. 1991 Sep 5;266(25):16376–16379. [PubMed] [Google Scholar]
  14. Hieter P., Mann C., Snyder M., Davis R. W. Mitotic stability of yeast chromosomes: a colony color assay that measures nondisjunction and chromosome loss. Cell. 1985 Feb;40(2):381–392. doi: 10.1016/0092-8674(85)90152-7. [DOI] [PubMed] [Google Scholar]
  15. Hill A., Bloom K. Genetic manipulation of centromere function. Mol Cell Biol. 1987 Jul;7(7):2397–2405. doi: 10.1128/mcb.7.7.2397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Holloway S. L., Glotzer M., King R. W., Murray A. W. Anaphase is initiated by proteolysis rather than by the inactivation of maturation-promoting factor. Cell. 1993 Jul 2;73(7):1393–1402. doi: 10.1016/0092-8674(93)90364-v. [DOI] [PubMed] [Google Scholar]
  17. Hoyt M. A., Totis L., Roberts B. T. S. cerevisiae genes required for cell cycle arrest in response to loss of microtubule function. Cell. 1991 Aug 9;66(3):507–517. doi: 10.1016/0092-8674(81)90014-3. [DOI] [PubMed] [Google Scholar]
  18. Koshland D., Rutledge L., Fitzgerald-Hayes M., Hartwell L. H. A genetic analysis of dicentric minichromosomes in Saccharomyces cerevisiae. Cell. 1987 Mar 13;48(5):801–812. doi: 10.1016/0092-8674(87)90077-8. [DOI] [PubMed] [Google Scholar]
  19. Kung A. L., Sherwood S. W., Schimke R. T. 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. doi: 10.1073/pnas.87.24.9553. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Li R., Murray A. W. Feedback control of mitosis in budding yeast. Cell. 1991 Aug 9;66(3):519–531. doi: 10.1016/0092-8674(81)90015-5. [DOI] [PubMed] [Google Scholar]
  21. Li X., Nicklas R. B. Mitotic forces control a cell-cycle checkpoint. Nature. 1995 Feb 16;373(6515):630–632. doi: 10.1038/373630a0. [DOI] [PubMed] [Google Scholar]
  22. Murray A. W., Schultes N. P., Szostak J. W. Chromosome length controls mitotic chromosome segregation in yeast. Cell. 1986 May 23;45(4):529–536. doi: 10.1016/0092-8674(86)90284-9. [DOI] [PubMed] [Google Scholar]
  23. Murray A. W., Szostak J. W. Chromosome segregation in mitosis and meiosis. Annu Rev Cell Biol. 1985;1:289–315. doi: 10.1146/annurev.cb.01.110185.001445. [DOI] [PubMed] [Google Scholar]
  24. Murray A. W., Szostak J. W. Construction and behavior of circularly permuted and telocentric chromosomes in Saccharomyces cerevisiae. Mol Cell Biol. 1986 Sep;6(9):3166–3172. doi: 10.1128/mcb.6.9.3166. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murray A. W., Szostak J. W. Construction of artificial chromosomes in yeast. Nature. 1983 Sep 15;305(5931):189–193. doi: 10.1038/305189a0. [DOI] [PubMed] [Google Scholar]
  26. Murray A. W., Szostak J. W. Pedigree analysis of plasmid segregation in yeast. Cell. 1983 Oct;34(3):961–970. doi: 10.1016/0092-8674(83)90553-6. [DOI] [PubMed] [Google Scholar]
  27. Murray A. W. The genetics of cell cycle checkpoints. Curr Opin Genet Dev. 1995 Feb;5(1):5–11. doi: 10.1016/s0959-437x(95)90046-2. [DOI] [PubMed] [Google Scholar]
  28. Murray A. Cell cycle checkpoints. Curr Opin Cell Biol. 1994 Dec;6(6):872–876. doi: 10.1016/0955-0674(94)90059-0. [DOI] [PubMed] [Google Scholar]
  29. Neff M. W., Burke D. J. A delay in the Saccharomyces cerevisiae cell cycle that is induced by a dicentric chromosome and dependent upon mitotic checkpoints. Mol Cell Biol. 1992 Sep;12(9):3857–3864. doi: 10.1128/mcb.12.9.3857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nicklas R. B., Koch C. A. Chromosome micromanipulation. 3. Spindle fiber tension and the reorientation of mal-oriented chromosomes. J Cell Biol. 1969 Oct;43(1):40–50. doi: 10.1083/jcb.43.1.40. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Nicklas R. B., Ward S. C., Gorbsky G. J. Kinetochore chemistry is sensitive to tension and may link mitotic forces to a cell cycle checkpoint. J Cell Biol. 1995 Aug;130(4):929–939. doi: 10.1083/jcb.130.4.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Raghuraman M. K., Brewer B. J., Fangman W. L. Activation of a yeast replication origin near a double-stranded DNA break. Genes Dev. 1994 Mar 1;8(5):554–562. doi: 10.1101/gad.8.5.554. [DOI] [PubMed] [Google Scholar]
  33. Rieder C. L., Alexander S. P. Kinetochores are transported poleward along a single astral microtubule during chromosome attachment to the spindle in newt lung cells. J Cell Biol. 1990 Jan;110(1):81–95. doi: 10.1083/jcb.110.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rieder C. L., Cole R. W., Khodjakov A., Sluder G. The checkpoint delaying anaphase in response to chromosome monoorientation is mediated by an inhibitory signal produced by unattached kinetochores. J Cell Biol. 1995 Aug;130(4):941–948. doi: 10.1083/jcb.130.4.941. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rieder C. L., Davison E. A., Jensen L. C., Cassimeris L., Salmon E. D. Oscillatory movements of monooriented chromosomes and their position relative to the spindle pole result from the ejection properties of the aster and half-spindle. J Cell Biol. 1986 Aug;103(2):581–591. doi: 10.1083/jcb.103.2.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rieder C. L., Palazzo R. E. Colcemid and the mitotic cycle. J Cell Sci. 1992 Jul;102(Pt 3):387–392. doi: 10.1242/jcs.102.3.387. [DOI] [PubMed] [Google Scholar]
  37. Rieder C. L., Salmon E. D. Motile kinetochores and polar ejection forces dictate chromosome position on the vertebrate mitotic spindle. J Cell Biol. 1994 Feb;124(3):223–233. doi: 10.1083/jcb.124.3.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rieder C. L., Schultz A., Cole R., Sluder G. Anaphase onset in vertebrate somatic cells is controlled by a checkpoint that monitors sister kinetochore attachment to the spindle. J Cell Biol. 1994 Dec;127(5):1301–1310. doi: 10.1083/jcb.127.5.1301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Roberts B. T., Farr K. A., Hoyt M. A. The Saccharomyces cerevisiae checkpoint gene BUB1 encodes a novel protein kinase. Mol Cell Biol. 1994 Dec;14(12):8282–8291. doi: 10.1128/mcb.14.12.8282. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Runge K. W., Wellinger R. J., Zakian V. A. Effects of excess centromeres and excess telomeres on chromosome loss rates. Mol Cell Biol. 1991 Jun;11(6):2919–2928. doi: 10.1128/mcb.11.6.2919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Sandell L. L., Zakian V. A. Loss of a yeast telomere: arrest, recovery, and chromosome loss. Cell. 1993 Nov 19;75(4):729–739. doi: 10.1016/0092-8674(93)90493-a. [DOI] [PubMed] [Google Scholar]
  42. Skibbens R. V., Skeen V. P., Salmon E. D. 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. doi: 10.1083/jcb.122.4.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Sluder G., Miller F. J., Spanjian K. The role of spindle microtubules in the timing of the cell cycle in echinoderm eggs. J Exp Zool. 1986 Jun;238(3):325–336. doi: 10.1002/jez.1402380307. [DOI] [PubMed] [Google Scholar]
  44. Sluder G., Miller F. J., Thompson E. A., Wolf D. E. Feedback control of the metaphase-anaphase transition in sea urchin zygotes: role of maloriented chromosomes. J Cell Biol. 1994 Jul;126(1):189–198. doi: 10.1083/jcb.126.1.189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Smith D. R., Smyth A. P., Moir D. T. Amplification of large artificial chromosomes. Proc Natl Acad Sci U S A. 1990 Nov;87(21):8242–8246. doi: 10.1073/pnas.87.21.8242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. 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. doi: 10.1073/pnas.89.19.8908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Tomkiel J., Cooke C. A., Saitoh H., Bernat R. L., Earnshaw W. C. 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. doi: 10.1083/jcb.125.3.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weinert T. A., Hartwell L. H. The RAD9 gene controls the cell cycle response to DNA damage in Saccharomyces cerevisiae. Science. 1988 Jul 15;241(4863):317–322. doi: 10.1126/science.3291120. [DOI] [PubMed] [Google Scholar]

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