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. 1988 Feb;8(2):754–763. doi: 10.1128/mcb.8.2.754

Structural organization and functional analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe.

B Fishel 1, H Amstutz 1, M Baum 1, J Carbon 1, L Clarke 1
PMCID: PMC363201  PMID: 2832735

Abstract

Centromeric DNA in the fission yeast Schizosaccharomyces pombe was isolated by chromosome walking and by field inversion gel electrophoretic fractionation of large genomic DNA restriction fragments. The centromere regions of the three chromosomes were contained on three SalI fragments (120 kilobases [kb], chromosome III; 90 kb, chromosome II; and 50 kb, chromosome I). Each fragment contained several repetitive DNA sequences, including repeat K (6.4 kb), repeat L (6.0 kb), and repeat B, that occurred only in the three centromere regions. On chromosome II, these repeats were organized into a 35-kb inverted repeat that included one copy of K and L in each arm of the repeat. Site-directed integration of a plasmid containing the yeast LEU2 gene into K repeats at each of the centromeres or integration of an intact K repeat into a chromosome arm had no effect on mitotic or meiotic centromere function. The centromeric repeat sequences were not transcribed and possessed many of the properties of constitutive heterochromatin. Thus, S. pombe is an excellent model system for studies on the role of repetitive sequence elements in centromere function.

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Selected References

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

  1. Amstutz H., Munz P., Heyer W. D., Leupoid U., Kohli J. Concerted evolution of tRNA genes: intergenic conversion among three unlinked serine tRNA genes in S. pombe. Cell. 1985 Apr;40(4):879–886. doi: 10.1016/0092-8674(85)90347-2. [DOI] [PubMed] [Google Scholar]
  2. Beach D. H., Klar A. J. Rearrangements of the transposable mating-type cassettes of fission yeast. EMBO J. 1984 Mar;3(3):603–610. doi: 10.1002/j.1460-2075.1984.tb01855.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Burke D. T., Carle G. F., Olson M. V. Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors. Science. 1987 May 15;236(4803):806–812. doi: 10.1126/science.3033825. [DOI] [PubMed] [Google Scholar]
  4. Butner K., Lo C. W. Modulation of tk expression in mouse pericentromeric heterochromatin. Mol Cell Biol. 1986 Dec;6(12):4440–4449. doi: 10.1128/mcb.6.12.4440. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carle G. F., Frank M., Olson M. V. Electrophoretic separations of large DNA molecules by periodic inversion of the electric field. Science. 1986 Apr 4;232(4746):65–68. doi: 10.1126/science.3952500. [DOI] [PubMed] [Google Scholar]
  6. Chan C. S., Tye B. K. Organization of DNA sequences and replication origins at yeast telomeres. Cell. 1983 Jun;33(2):563–573. doi: 10.1016/0092-8674(83)90437-3. [DOI] [PubMed] [Google Scholar]
  7. Clarke L., Amstutz H., Fishel B., Carbon J. Analysis of centromeric DNA in the fission yeast Schizosaccharomyces pombe. Proc Natl Acad Sci U S A. 1986 Nov;83(21):8253–8257. doi: 10.1073/pnas.83.21.8253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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]
  9. Clarke L., Carbon J. The structure and function of yeast centromeres. Annu Rev Genet. 1985;19:29–55. doi: 10.1146/annurev.ge.19.120185.000333. [DOI] [PubMed] [Google Scholar]
  10. Collins J. Instability of palindromic DNA in Escherichia coli. Cold Spring Harb Symp Quant Biol. 1981;45(Pt 1):409–416. doi: 10.1101/sqb.1981.045.01.055. [DOI] [PubMed] [Google Scholar]
  11. Flores da Cunha M. Mitotic mapping of Schizosaccharomyces pombe. Genet Res. 1970 Oct 2;16(2):127–144. doi: 10.1017/s0016672300002366. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Heyer W. D., Sipiczki M., Kohli J. Replicating plasmids in Schizosaccharomyces pombe: improvement of symmetric segregation by a new genetic element. Mol Cell Biol. 1986 Jan;6(1):80–89. doi: 10.1128/mcb.6.1.80. [DOI] [PMC free article] [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. Hsiao C. L., Carbon J. Direct selection procedure for the isolation of functional centromeric DNA. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3760–3764. doi: 10.1073/pnas.78.6.3760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ish-Horowicz D., Burke J. F. Rapid and efficient cosmid cloning. Nucleic Acids Res. 1981 Jul 10;9(13):2989–2998. doi: 10.1093/nar/9.13.2989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Ito H., Fukuda Y., Murata K., Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. doi: 10.1128/jb.153.1.163-168.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kohli J., Hottinger H., Munz P., Strauss A., Thuriaux P. Genetic Mapping in SCHIZOSACCHAROMYCES POMBE by Mitotic and Meiotic Analysis and Induced Haploidization. Genetics. 1977 Nov;87(3):471–489. doi: 10.1093/genetics/87.3.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. LEWIS E. B. The phenomenon of position effect. Adv Genet. 1950;3:73–115. doi: 10.1016/s0065-2660(08)60083-8. [DOI] [PubMed] [Google Scholar]
  20. Leach D. R., Stahl F. W. Viability of lambda phages carrying a perfect palindrome in the absence of recombination nucleases. 1983 Sep 29-Oct 5Nature. 305(5933):448–451. doi: 10.1038/305448a0. [DOI] [PubMed] [Google Scholar]
  21. Nakaseko Y., Adachi Y., Funahashi S., Niwa O., Yanagida M. Chromosome walking shows a highly homologous repetitive sequence present in all the centromere regions of fission yeast. EMBO J. 1986 May;5(5):1011–1021. doi: 10.1002/j.1460-2075.1986.tb04316.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nakaseko Y., Kinoshita N., Yanagida M. A novel sequence common to the centromere regions of Schizosaccharomyces pombe chromosomes. Nucleic Acids Res. 1987 Jun 25;15(12):4705–4715. doi: 10.1093/nar/15.12.4705. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Orr-Weaver T. L., Szostak J. W., Rothstein R. J. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. doi: 10.1073/pnas.78.10.6354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Robinow C. F. The Number of Chromosomes in SCHIZOSACCHAROMYCES POMBE: Light Microscopy of Stained Preparations. Genetics. 1977 Nov;87(3):491–497. doi: 10.1093/genetics/87.3.491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rushlow C. A., Bender W., Chovnick A. Studies on the mechanism of heterochromatic position effect at the rosy locus of Drosophila melanogaster. Genetics. 1984 Nov;108(3):603–615. doi: 10.1093/genetics/108.3.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Singer M. F. Highly repeated sequences in mammalian genomes. Int Rev Cytol. 1982;76:67–112. doi: 10.1016/s0074-7696(08)61789-1. [DOI] [PubMed] [Google Scholar]
  27. Steensma H. Y., Crowley J. C., Kaback D. B. Molecular cloning of chromosome I DNA from Saccharomyces cerevisiae: isolation and analysis of the CEN1-ADE1-CDC15 region. Mol Cell Biol. 1987 Jan;7(1):410–419. doi: 10.1128/mcb.7.1.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Tsao S. G., Brunk C. F., Pearlman R. E. Hybridization of nucleic acids directly in agarose gels. Anal Biochem. 1983 Jun;131(2):365–372. doi: 10.1016/0003-2697(83)90185-9. [DOI] [PubMed] [Google Scholar]
  29. Umesono K., Toda T., Hayashi S., Yanagida M. Cell division cycle genes nda2 and nda3 of the fission yeast Schizosaccharomyces pombe control microtubular organization and sensitivity to anti-mitotic benzimidazole compounds. J Mol Biol. 1983 Aug 5;168(2):271–284. doi: 10.1016/s0022-2836(83)80018-7. [DOI] [PubMed] [Google Scholar]
  30. Wyman A. R., Wolfe L. B., Botstein D. Propagation of some human DNA sequences in bacteriophage lambda vectors requires mutant Escherichia coli hosts. Proc Natl Acad Sci U S A. 1985 May;82(9):2880–2884. doi: 10.1073/pnas.82.9.2880. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yeh E., Carbon J., Bloom K. Tightly centromere-linked gene (SPO15) essential for meiosis in the yeast Saccharomyces cerevisiae. Mol Cell Biol. 1986 Jan;6(1):158–167. doi: 10.1128/mcb.6.1.158. [DOI] [PMC free article] [PubMed] [Google Scholar]

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