Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1991 Jun;11(6):2919–2928. doi: 10.1128/mcb.11.6.2919

Effects of excess centromeres and excess telomeres on chromosome loss rates.

K W Runge 1, R J Wellinger 1, V A Zakian 1
PMCID: PMC360116  PMID: 2038311

Abstract

The linear chromosomes of eukaryotes contain specialized structures to ensure their faithful replication and segregation to daughter cells. Two of these structures, centromeres and telomeres, are limited, respectively, to one and two copies per chromosome. It is possible that the proteins that interact with centromere and telomere DNA sequences are present in limiting amounts and could be competed away from the chromosomal copies of these elements by additional copies introduced on plasmids. We have introduced excess centromeres and telomeres into Saccharomyces cerevisiae and quantitated their effects on the rates of loss of chromosome III and chromosome VII by fluctuation analysis. We show that (i) 600 new telomeres have no effect on chromosome loss; (ii) an average of 25 extra centromere DNA sequences increase the rate of chromosome III loss from 0.4 x 10(-4) events per cell division to 1.3 x 10(-3) events per cell division; (iii) centromere DNA (CEN) sequences on circular vectors destabilize chromosomes more effectively than do CEN sequences on 15-kb linear vectors, and transcribed CEN sequences have no effect on chromosome stability. We discuss the different effects of extra centromere and telomere DNA sequences on chromosome stability in terms of how the cell recognizes these two chromosomal structures.

Full text

PDF
2919

Selected References

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

  1. Alani E., Cao L., Kleckner N. A method for gene disruption that allows repeated use of URA3 selection in the construction of multiply disrupted yeast strains. Genetics. 1987 Aug;116(4):541–545. doi: 10.1534/genetics.112.541.test. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baker R. E., Fitzgerald-Hayes M., O'Brien T. C. Purification of the yeast centromere binding protein CP1 and a mutational analysis of its binding site. J Biol Chem. 1989 Jun 25;264(18):10843–10850. [PubMed] [Google Scholar]
  3. Baker R. E., Masison D. C. Isolation of the gene encoding the Saccharomyces cerevisiae centromere-binding protein CP1. Mol Cell Biol. 1990 Jun;10(6):2458–2467. doi: 10.1128/mcb.10.6.2458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biessmann H., Carter S. B., Mason J. M. Chromosome ends in Drosophila without telomeric DNA sequences. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1758–1761. doi: 10.1073/pnas.87.5.1758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bloom K. S., Amaya E., Carbon J., Clarke L., Hill A., Yeh E. Chromatin conformation of yeast centromeres. J Cell Biol. 1984 Nov;99(5):1559–1568. doi: 10.1083/jcb.99.5.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bloom K. S., Carbon J. Yeast centromere DNA is in a unique and highly ordered structure in chromosomes and small circular minichromosomes. Cell. 1982 Jun;29(2):305–317. doi: 10.1016/0092-8674(82)90147-7. [DOI] [PubMed] [Google Scholar]
  7. Bram R. J., Kornberg R. D. Isolation of a Saccharomyces cerevisiae centromere DNA-binding protein, its human homolog, and its possible role as a transcription factor. Mol Cell Biol. 1987 Jan;7(1):403–409. doi: 10.1128/mcb.7.1.403. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buchman A. R., Lue N. F., Kornberg R. D. Connections between transcriptional activators, silencers, and telomeres as revealed by functional analysis of a yeast DNA-binding protein. Mol Cell Biol. 1988 Dec;8(12):5086–5099. doi: 10.1128/mcb.8.12.5086. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Cai M., Davis R. W. Yeast centromere binding protein CBF1, of the helix-loop-helix protein family, is required for chromosome stability and methionine prototrophy. Cell. 1990 May 4;61(3):437–446. doi: 10.1016/0092-8674(90)90525-j. [DOI] [PubMed] [Google Scholar]
  10. Campbell D. A., Fogel S. Association of chromosome loss with centromere-adjacent mitotic recombination in a yeast disomic haploid. Genetics. 1977 Apr;85(4):573–585. doi: 10.1093/genetics/85.4.573. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Campbell D. A., Fogel S., Lusnak K. Mitotic chromosome loss in a disomic haploid of Saccharomyces cerevisiae. Genetics. 1975 Mar;79(3):383–396. doi: 10.1093/genetics/79.3.383. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Carle G. F., Olson M. V. An electrophoretic karyotype for yeast. Proc Natl Acad Sci U S A. 1985 Jun;82(11):3756–3760. doi: 10.1073/pnas.82.11.3756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Conrad M. N., Wright J. H., Wolf A. J., Zakian V. A. RAP1 protein interacts with yeast telomeres in vivo: overproduction alters telomere structure and decreases chromosome stability. Cell. 1990 Nov 16;63(4):739–750. doi: 10.1016/0092-8674(90)90140-a. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. Dutcher S. K., Hartwell L. H. Test for temporal or spatial restrictions in gene product function during the cell division cycle. Mol Cell Biol. 1983 Jul;3(7):1255–1265. doi: 10.1128/mcb.3.7.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dutcher S. K. Internuclear transfer of genetic information in kar1-1/KAR1 heterokaryons in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Mar;1(3):245–253. doi: 10.1128/mcb.1.3.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Esposito M. S., Maleas D. T., Bjornstad K. A., Holbrook L. L. The REC46 gene of Saccharomyces cerevisiae controls mitotic chromosomal stability, recombination and sporulation: cell-type and life cycle stage-specific expression of the rec46-1 mutation. Curr Genet. 1986;10(6):425–433. doi: 10.1007/BF00419869. [DOI] [PubMed] [Google Scholar]
  19. Fitzgerald-Hayes M. Yeast centromeres. Yeast. 1987 Sep;3(3):187–200. doi: 10.1002/yea.320030306. [DOI] [PubMed] [Google Scholar]
  20. Haber J. E., Thorburn P. C. Healing of broken linear dicentric chromosomes in yeast. Genetics. 1984 Feb;106(2):207–226. doi: 10.1093/genetics/106.2.207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Haber J. E., Thorburn P. C., Rogers D. Meiotic and mitotic behavior of dicentric chromosomes in Saccharomyces cerevisiae. Genetics. 1984 Feb;106(2):185–205. doi: 10.1093/genetics/106.2.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hartwell L. H., Smith D. Altered fidelity of mitotic chromosome transmission in cell cycle mutants of S. cerevisiae. Genetics. 1985 Jul;110(3):381–395. doi: 10.1093/genetics/110.3.381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hegemann J. H., Shero J. H., Cottarel G., Philippsen P., Hieter P. Mutational analysis of centromere DNA from chromosome VI of Saccharomyces cerevisiae. Mol Cell Biol. 1988 Jun;8(6):2523–2535. doi: 10.1128/mcb.8.6.2523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Hoyt M. A., Stearns T., Botstein D. Chromosome instability mutants of Saccharomyces cerevisiae that are defective in microtubule-mediated processes. Mol Cell Biol. 1990 Jan;10(1):223–234. doi: 10.1128/mcb.10.1.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Huffaker T. C., Thomas J. H., Botstein D. Diverse effects of beta-tubulin mutations on microtubule formation and function. J Cell Biol. 1988 Jun;106(6):1997–2010. doi: 10.1083/jcb.106.6.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. 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]
  28. Larionov V. L., Kouprina N. Y., Strunnikov A. V., Vlasov A. V. A direct selection procedure for isolating yeast mutants with an impaired segregation of artificial minichromosomes. Curr Genet. 1989 Jan;15(1):17–25. doi: 10.1007/BF00445747. [DOI] [PubMed] [Google Scholar]
  29. Levis R. W. Viable deletions of a telomere from a Drosophila chromosome. Cell. 1989 Aug 25;58(4):791–801. doi: 10.1016/0092-8674(89)90112-8. [DOI] [PubMed] [Google Scholar]
  30. Lundblad V., Szostak J. W. A mutant with a defect in telomere elongation leads to senescence in yeast. Cell. 1989 May 19;57(4):633–643. doi: 10.1016/0092-8674(89)90132-3. [DOI] [PubMed] [Google Scholar]
  31. Lustig A. J., Kurtz S., Shore D. Involvement of the silencer and UAS binding protein RAP1 in regulation of telomere length. Science. 1990 Oct 26;250(4980):549–553. doi: 10.1126/science.2237406. [DOI] [PubMed] [Google Scholar]
  32. Mann C., Davis R. W. Instability of dicentric plasmids in yeast. Proc Natl Acad Sci U S A. 1983 Jan;80(1):228–232. doi: 10.1073/pnas.80.1.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. McClintock B. The Behavior in Successive Nuclear Divisions of a Chromosome Broken at Meiosis. Proc Natl Acad Sci U S A. 1939 Aug;25(8):405–416. doi: 10.1073/pnas.25.8.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Meeks-Wagner D., Hartwell L. H. Normal stoichiometry of histone dimer sets is necessary for high fidelity of mitotic chromosome transmission. Cell. 1986 Jan 17;44(1):43–52. doi: 10.1016/0092-8674(86)90483-6. [DOI] [PubMed] [Google Scholar]
  35. Meeks-Wagner D., Wood J. S., Garvik B., Hartwell L. H. Isolation of two genes that affect mitotic chromosome transmission in S. cerevisiae. Cell. 1986 Jan 17;44(1):53–63. doi: 10.1016/0092-8674(86)90484-8. [DOI] [PubMed] [Google Scholar]
  36. 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]
  37. 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]
  38. 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]
  39. 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]
  40. Newlon C. S. Yeast chromosome replication and segregation. Microbiol Rev. 1988 Dec;52(4):568–601. doi: 10.1128/mr.52.4.568-601.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Oertel W., Mayer M. Structure and mitotic stability of minichromosomes originating in yeast cells transformed with tandem dimers of CEN11 plasmids. Mol Gen Genet. 1984;195(1-2):300–307. doi: 10.1007/BF00332763. [DOI] [PubMed] [Google Scholar]
  42. Pluta A. F., Zakian V. A. Recombination occurs during telomere formation in yeast. Nature. 1989 Feb 2;337(6206):429–433. doi: 10.1038/337429a0. [DOI] [PubMed] [Google Scholar]
  43. Rose M. D. Isolation of genes by complementation in yeast. Methods Enzymol. 1987;152:481–504. doi: 10.1016/0076-6879(87)52056-0. [DOI] [PubMed] [Google Scholar]
  44. Rose M., Grisafi P., Botstein D. Structure and function of the yeast URA3 gene: expression in Escherichia coli. Gene. 1984 Jul-Aug;29(1-2):113–124. doi: 10.1016/0378-1119(84)90172-0. [DOI] [PubMed] [Google Scholar]
  45. Runge K. W., Zakian V. A. Introduction of extra telomeric DNA sequences into Saccharomyces cerevisiae results in telomere elongation. Mol Cell Biol. 1989 Apr;9(4):1488–1497. doi: 10.1128/mcb.9.4.1488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Runge K. W., Zakian V. A. Properties of the transcriptional enhancer in Saccharomyces cerevisiae telomeres. Nucleic Acids Res. 1990 Apr 11;18(7):1783–1787. doi: 10.1093/nar/18.7.1783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Saunders M., Fitzgerald-Hayes M., Bloom K. Chromatin structure of altered yeast centromeres. Proc Natl Acad Sci U S A. 1988 Jan;85(1):175–179. doi: 10.1073/pnas.85.1.175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Spencer F., Gerring S. L., Connelly C., Hieter P. Mitotic chromosome transmission fidelity mutants in Saccharomyces cerevisiae. Genetics. 1990 Feb;124(2):237–249. doi: 10.1093/genetics/124.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Stinchcomb D. T., Mann C., Davis R. W. Centromeric DNA from Saccharomyces cerevisiae. J Mol Biol. 1982 Jun 25;158(2):157–190. doi: 10.1016/0022-2836(82)90427-2. [DOI] [PubMed] [Google Scholar]
  50. Surosky R. T., Tye B. K. Construction of telocentric chromosomes in Saccharomyces cerevisiae. Proc Natl Acad Sci U S A. 1985 Apr;82(7):2106–2110. doi: 10.1073/pnas.82.7.2106. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Surosky R. T., Tye B. K. Resolution of dicentric chromosomes by Ty-mediated recombination in yeast. Genetics. 1985 Jul;110(3):397–419. doi: 10.1093/genetics/110.3.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Tschumper G., Carbon J. Sequence of a yeast DNA fragment containing a chromosomal replicator and the TRP1 gene. Gene. 1980 Jul;10(2):157–166. doi: 10.1016/0378-1119(80)90133-x. [DOI] [PubMed] [Google Scholar]
  53. Vollrath D., Davis R. W. Resolution of DNA molecules greater than 5 megabases by contour-clamped homogeneous electric fields. Nucleic Acids Res. 1987 Oct 12;15(19):7865–7876. doi: 10.1093/nar/15.19.7865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Wellinger R. J., Zakian V. A. Lack of positional requirements for autonomously replicating sequence elements on artificial yeast chromosomes. Proc Natl Acad Sci U S A. 1989 Feb;86(3):973–977. doi: 10.1073/pnas.86.3.973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Zakian V. A., Blanton H. M., Wetzel L., Dani G. M. Size threshold for Saccharomyces cerevisiae chromosomes: generation of telocentric chromosomes from an unstable minichromosome. Mol Cell Biol. 1986 Mar;6(3):925–932. doi: 10.1128/mcb.6.3.925. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Zakian V. A., Kupfer D. M. Replication and segregation of an unstable plasmid in yeast. Plasmid. 1982 Jul;8(1):15–28. doi: 10.1016/0147-619x(82)90037-3. [DOI] [PubMed] [Google Scholar]
  57. Zakian V. A. Structure and function of telomeres. Annu Rev Genet. 1989;23:579–604. doi: 10.1146/annurev.ge.23.120189.003051. [DOI] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES