Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 2001 Dec;159(4):1501–1509. doi: 10.1093/genetics/159.4.1501

Spontaneous chromosome loss in Saccharomyces cerevisiae is suppressed by DNA damage checkpoint functions.

H L Klein 1
PMCID: PMC1461919  PMID: 11779792

Abstract

Genomic instability is one of the hallmarks of cancer cells and is often the causative factor in revealing recessive gene mutations that progress cells along the pathway to unregulated growth. Genomic instability can take many forms, including aneuploidy and changes in chromosome structure. Chromosome loss, loss and reduplication, and deletions are the majority events that result in loss of heterozygosity (LOH). Defective DNA replication, repair, and recombination can significantly increase the frequency of spontaneous genomic instability. Recently, DNA damage checkpoint functions that operate during the S-phase checkpoint have been shown to suppress spontaneous chromosome rearrangements in haploid yeast strains. To further study the role of DNA damage checkpoint functions in genomic stability, we have determined chromosome loss in DNA damage checkpoint-deficient yeast strains. We have found that the DNA damage checkpoints are essential for preserving the normal chromosome number and act synergistically with homologous recombination functions to ensure that chromosomes are segregated correctly to daughter cells. Failure of either of these processes increases LOH events. However, loss of the G2/M checkpoint does not result in an increase in chromosome loss, suggesting that it is the various S-phase DNA damage checkpoints that suppress chromosome loss. The mec1 checkpoint function mutant, defective in the yeast ATR homolog, results in increased recombination through a process that is distinct from that operative in wild-type cells.

Full Text

The Full Text of this article is available as a PDF (121.8 KB).

Selected References

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

  1. Bashkirov V. I., King J. S., Bashkirova E. V., Schmuckli-Maurer J., Heyer W. D. DNA repair protein Rad55 is a terminal substrate of the DNA damage checkpoints. Mol Cell Biol. 2000 Jun;20(12):4393–4404. doi: 10.1128/mcb.20.12.4393-4404.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boulton S. J., Jackson S. P. Saccharomyces cerevisiae Ku70 potentiates illegitimate DNA double-strand break repair and serves as a barrier to error-prone DNA repair pathways. EMBO J. 1996 Sep 16;15(18):5093–5103. [PMC free article] [PubMed] [Google Scholar]
  3. Brown E. J., Baltimore D. ATR disruption leads to chromosomal fragmentation and early embryonic lethality. Genes Dev. 2000 Feb 15;14(4):397–402. [PMC free article] [PubMed] [Google Scholar]
  4. Chen C., Kolodner R. D. Gross chromosomal rearrangements in Saccharomyces cerevisiae replication and recombination defective mutants. Nat Genet. 1999 Sep;23(1):81–85. doi: 10.1038/12687. [DOI] [PubMed] [Google Scholar]
  5. Clarke D. J., Segal M., Mondésert G., Reed S. I. The Pds1 anaphase inhibitor and Mec1 kinase define distinct checkpoints coupling S phase with mitosis in budding yeast. Curr Biol. 1999 Apr 8;9(7):365–368. doi: 10.1016/s0960-9822(99)80163-8. [DOI] [PubMed] [Google Scholar]
  6. Cohen-Fix O., Koshland D. The anaphase inhibitor of Saccharomyces cerevisiae Pds1p is a target of the DNA damage checkpoint pathway. Proc Natl Acad Sci U S A. 1997 Dec 23;94(26):14361–14366. doi: 10.1073/pnas.94.26.14361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Datta A., Schmeits J. L., Amin N. S., Lau P. J., Myung K., Kolodner R. D. Checkpoint-dependent activation of mutagenic repair in Saccharomyces cerevisiae pol3-01 mutants. Mol Cell. 2000 Sep;6(3):593–603. doi: 10.1016/s1097-2765(00)00058-7. [DOI] [PubMed] [Google Scholar]
  8. Difilippantonio M. J., Zhu J., Chen H. T., Meffre E., Nussenzweig M. C., Max E. E., Ried T., Nussenzweig A. DNA repair protein Ku80 suppresses chromosomal aberrations and malignant transformation. Nature. 2000 Mar 30;404(6777):510–514. doi: 10.1038/35006670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fasullo M., Bennett T., AhChing P., Koudelik J. The Saccharomyces cerevisiae RAD9 checkpoint reduces the DNA damage-associated stimulation of directed translocations. Mol Cell Biol. 1998 Mar;18(3):1190–1200. doi: 10.1128/mcb.18.3.1190. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ferguson D. O., Sekiguchi J. M., Chang S., Frank K. M., Gao Y., DePinho R. A., Alt F. W. The nonhomologous end-joining pathway of DNA repair is required for genomic stability and the suppression of translocations. Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6630–6633. doi: 10.1073/pnas.110152897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Foiani M., Ferrari M., Liberi G., Lopes M., Lucca C., Marini F., Pellicioli A., Muzi Falconi M., Plevani P. S-phase DNA damage checkpoint in budding yeast. Biol Chem. 1998 Aug-Sep;379(8-9):1019–1023. doi: 10.1515/bchm.1998.379.8-9.1019. [DOI] [PubMed] [Google Scholar]
  12. Frei C., Gasser S. M. The yeast Sgs1p helicase acts upstream of Rad53p in the DNA replication checkpoint and colocalizes with Rad53p in S-phase-specific foci. Genes Dev. 2000 Jan 1;14(1):81–96. [PMC free article] [PubMed] [Google Scholar]
  13. Friis J., Roman H. The effect of the mating-type alleles on intragenic recombination in yeast. Genetics. 1968 May;59(1):33–36. doi: 10.1093/genetics/59.1.33. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Galgoczy D. J., Toczyski D. P. Checkpoint adaptation precedes spontaneous and damage-induced genomic instability in yeast. Mol Cell Biol. 2001 Mar;21(5):1710–1718. doi: 10.1128/MCB.21.5.1710-1718.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grushcow J. M., Holzen T. M., Park K. J., Weinert T., Lichten M., Bishop D. K. Saccharomyces cerevisiae checkpoint genes MEC1, RAD17 and RAD24 are required for normal meiotic recombination partner choice. Genetics. 1999 Oct;153(2):607–620. doi: 10.1093/genetics/153.2.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Haber J. E. Recombination: a frank view of exchanges and vice versa. Curr Opin Cell Biol. 2000 Jun;12(3):286–292. doi: 10.1016/s0955-0674(00)00090-9. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Hartwell L. Defects in a cell cycle checkpoint may be responsible for the genomic instability of cancer cells. Cell. 1992 Nov 13;71(4):543–546. doi: 10.1016/0092-8674(92)90586-2. [DOI] [PubMed] [Google Scholar]
  19. Hartwell L., Weinert T., Kadyk L., Garvik B. Cell cycle checkpoints, genomic integrity, and cancer. Cold Spring Harb Symp Quant Biol. 1994;59:259–263. doi: 10.1101/sqb.1994.059.01.030. [DOI] [PubMed] [Google Scholar]
  20. Hiraoka M., Watanabe K., Umezu K., Maki H. Spontaneous loss of heterozygosity in diploid Saccharomyces cerevisiae cells. Genetics. 2000 Dec;156(4):1531–1548. doi: 10.1093/genetics/156.4.1531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kadyk L. C., Hartwell L. H. Sister chromatids are preferred over homologs as substrates for recombinational repair in Saccharomyces cerevisiae. Genetics. 1992 Oct;132(2):387–402. doi: 10.1093/genetics/132.2.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Karanjawala Z. E., Grawunder U., Hsieh C. L., Lieber M. R. The nonhomologous DNA end joining pathway is important for chromosome stability in primary fibroblasts. Curr Biol. 1999 Dec 16;9(24):1501–1504. doi: 10.1016/s0960-9822(00)80123-2. [DOI] [PubMed] [Google Scholar]
  23. Karran P. DNA double strand break repair in mammalian cells. Curr Opin Genet Dev. 2000 Apr;10(2):144–150. doi: 10.1016/s0959-437x(00)00069-1. [DOI] [PubMed] [Google Scholar]
  24. Kato R., Ogawa H. An essential gene, ESR1, is required for mitotic cell growth, DNA repair and meiotic recombination in Saccharomyces cerevisiae. Nucleic Acids Res. 1994 Aug 11;22(15):3104–3112. doi: 10.1093/nar/22.15.3104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Kinzler K. W., Vogelstein B. Landscaping the cancer terrain. Science. 1998 May 15;280(5366):1036–1037. doi: 10.1126/science.280.5366.1036. [DOI] [PubMed] [Google Scholar]
  26. Lengauer C., Kinzler K. W., Vogelstein B. Genetic instabilities in human cancers. Nature. 1998 Dec 17;396(6712):643–649. doi: 10.1038/25292. [DOI] [PubMed] [Google Scholar]
  27. Malkova A., Ivanov E. L., Haber J. E. Double-strand break repair in the absence of RAD51 in yeast: a possible role for break-induced DNA replication. Proc Natl Acad Sci U S A. 1996 Jul 9;93(14):7131–7136. doi: 10.1073/pnas.93.14.7131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Malone R. E., Ward T., Lin S., Waring J. The RAD50 gene, a member of the double strand break repair epistasis group, is not required for spontaneous mitotic recombination in yeast. Curr Genet. 1990 Aug;18(2):111–116. doi: 10.1007/BF00312598. [DOI] [PubMed] [Google Scholar]
  29. Myung K., Datta A., Kolodner R. D. Suppression of spontaneous chromosomal rearrangements by S phase checkpoint functions in Saccharomyces cerevisiae. Cell. 2001 Feb 9;104(3):397–408. doi: 10.1016/s0092-8674(01)00227-6. [DOI] [PubMed] [Google Scholar]
  30. Paulovich A. G., Armour C. D., Hartwell L. H. The Saccharomyces cerevisiae RAD9, RAD17, RAD24 and MEC3 genes are required for tolerating irreparable, ultraviolet-induced DNA damage. Genetics. 1998 Sep;150(1):75–93. doi: 10.1093/genetics/150.1.75. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Petukhova G., Van Komen S., Vergano S., Klein H., Sung P. Yeast Rad54 promotes Rad51-dependent homologous DNA pairing via ATP hydrolysis-driven change in DNA double helix conformation. J Biol Chem. 1999 Oct 8;274(41):29453–29462. doi: 10.1074/jbc.274.41.29453. [DOI] [PubMed] [Google Scholar]
  32. Point D., Rodriguez J., Ferrante B., Brugère J. Cancers du voile du palais. Résultats de la chirurgie de rattrapage. Ann Otolaryngol Chir Cervicofac. 1987;104(6):395–397. [PubMed] [Google Scholar]
  33. Rattray A. J., Symington L. S. Multiple pathways for homologous recombination in Saccharomyces cerevisiae. Genetics. 1995 Jan;139(1):45–56. doi: 10.1093/genetics/139.1.45. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rattray A. J., Symington L. S. Use of a chromosomal inverted repeat to demonstrate that the RAD51 and RAD52 genes of Saccharomyces cerevisiae have different roles in mitotic recombination. Genetics. 1994 Nov;138(3):587–595. doi: 10.1093/genetics/138.3.587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Sanchez Y., Bachant J., Wang H., Hu F., Liu D., Tetzlaff M., Elledge S. J. Control of the DNA damage checkpoint by chk1 and rad53 protein kinases through distinct mechanisms. Science. 1999 Nov 5;286(5442):1166–1171. doi: 10.1126/science.286.5442.1166. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. Signon L., Malkova A., Naylor M. L., Klein H., Haber J. E. Genetic requirements for RAD51- and RAD54-independent break-induced replication repair of a chromosomal double-strand break. Mol Cell Biol. 2001 Mar;21(6):2048–2056. doi: 10.1128/MCB.21.6.2048-2056.2001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Thompson D. A., Stahl F. W. Genetic control of recombination partner preference in yeast meiosis. Isolation and characterization of mutants elevated for meiotic unequal sister-chromatid recombination. Genetics. 1999 Oct;153(2):621–641. doi: 10.1093/genetics/153.2.621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Weinert T. A., Hartwell L. H. Characterization of RAD9 of Saccharomyces cerevisiae and evidence that its function acts posttranslationally in cell cycle arrest after DNA damage. Mol Cell Biol. 1990 Dec;10(12):6554–6564. doi: 10.1128/mcb.10.12.6554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Weinert T. Yeast checkpoint controls and relevance to cancer. Cancer Surv. 1997;29:109–132. [PubMed] [Google Scholar]
  41. Yamamoto A., Guacci V., Koshland D. Pds1p, an inhibitor of anaphase in budding yeast, plays a critical role in the APC and checkpoint pathway(s). J Cell Biol. 1996 Apr;133(1):99–110. doi: 10.1083/jcb.133.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. de Klein A., Muijtjens M., van Os R., Verhoeven Y., Smit B., Carr A. M., Lehmann A. R., Hoeijmakers J. H. Targeted disruption of the cell-cycle checkpoint gene ATR leads to early embryonic lethality in mice. Curr Biol. 2000 Apr 20;10(8):479–482. doi: 10.1016/s0960-9822(00)00447-4. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES