Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Double–strand breaks on YACs during yeast meiosis may reflect meiotic recombination in the human genome

Nature Genetics volume 13, pages 481–484 (1996)Cite this article

Abstract

Meiotic recombination in the yeast Saccharomyces cerevisiae is initiated at double-strand breaks (DSBs), which occur preferentially at specific locations1–3. Genetically mapped regions of elevated meiotic recombination (‘hotspots’) coincide with meioticDSB sites, which can be identified on chromosome blots of meiotic DNA (refs 4,5; S.K. et al., manuscript submitted). The morphology of yeast artificial chromosomes (YACs) containing human DNA during the pachytene stage of meiosis resembles that of native yeast chromosomes6. Homologous YAC pairs segregate faithfully and recombine at the high rates characteristic of S. cerevisiae7 (vs. ∼0.4 cM/kb in S. cerevisiae versus ∼10−3 cM/kb in humans). We have examined a variety of YACs carrying human DNA inserts for double-strand breakage during yeast meiosis. Each YAC has a characteristic set of meiotic DSB sites, as do yeast chromosomes (S.K. et al., manuscript submitted). We show that the positions of the DSB sites in the YACs depend on the human-derived DNA in the clones. The degree of double-strand breakage in yeast meiosis of the YACs in our study appears to reflect the degree of meiotic recombination in humans.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Kleckner, N., Padmore, R. & Bishop, O.K. Meiotic chromosome metabolism: One view. Cold Spring Harb. Symp. Quant. Biol. 56, 729–743 (1991).

    Article  CAS  Google Scholar 

  2. Klein, S., Sherman, A. & Simchen, G. Regulation of meiosis and sporulation in Saccharomyces cerevisiae in Growth,. Differentiation and Sexuality, (eds Wessels, J.G.H. & Meinhardt, F.) 235–250 (Springer-Verlag, Berlin Heidelberg, 1994)

    Chapter  Google Scholar 

  3. Petes, T.D., Malone, R.E. & Symington, L.S. Recombination in yeast in The molecular and cellular biology of the yeast Saccharomyces: Genome dynamics, protein synthesis, and energetics. (eds Broach, J. R., Pringle, J.R. & Jones, E.W.) 407–521 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1991)

    Google Scholar 

  4. Zenvirth, D. et al. Multiple sites for double-strand breaks in whole meiotic chromosomes of Saccharomyces cerevisiae. EMBO J. 11, 3441–3447 (1992).

    Article  CAS  Google Scholar 

  5. Game, J.C. Pulsed-field gel analysis of the pattern of DMA double-strand breaks in the Saccharomyces genome during meiosis. Devl. Genet. 13, 485–497 (1992).

    Article  CAS  Google Scholar 

  6. Loidl, J., Scherthan, H., Den Dunnen, J.T. & Klein, F. Morphology of a human-derived YAC in yeast meiosis. Chromosoma 104, 183–188 (1995).

    Article  CAS  Google Scholar 

  7. Sears, D.D., Hegemann, J.H. & Hieter, P. Meiotic recombination and segregation of human derived artificial chromosomes in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 89, 5296–5300 (1992).

    Article  CAS  Google Scholar 

  8. den Dunnen,J.T et al. Reconstruction of the 2.4 Mb human DMD-gene by homologous YAC recombination. Hum. Mol. Genet. 1, 19–28 (1992).

    Article  CAS  Google Scholar 

  9. Monaco, A.P., Walker, A.P., Millwood, I., Larin, Z. & Lehrach, H. A yeast artificial chromosome contig containing the complete Duchenne muscular dystrophy gene. Genomics 12, 465–473 (1992).

    Article  CAS  Google Scholar 

  10. Henke, A., Fischer, C. & Rappold, G.A. Genetic map of the human pseudoautosomal region reveals a high rate of recombination in female meiosis at the Xp telomere. Genomics 18, 478–485 (1993).

    Article  CAS  Google Scholar 

  11. Ried, K. et al. Characterization of a YAC contig spanning the pseudoautosomal region. Genomics 29, 787–792 (1995).

    Article  CAS  Google Scholar 

  12. Chumakov, I. et al. Continuum of overlapping clones spanning the entire human chromosome 21q. Nature 359, 380–387 (1992).

    Article  CAS  Google Scholar 

  13. Treco, D., Thomas, B. & Arnheim, N. Recombination hot spot in the human β-globin gene cluster: Meiotic recombination of human DMA fragments in Saccharomyces cerevisiae. Mol. Cell. Biol. 5, 2029–2098 (1985).

    Article  CAS  Google Scholar 

  14. Ross, L.O., Treco, D., Nicolas, A., Szostak, J.W. & Dawson, D. Meiotic recombination on artificial chromosomes in yeast. Genetics 131, 541–550 (1992).

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Sun, H., Treco, D., Schultes, N.P. & Szostak, J.W. Double-strand breaks at an initiation site for meiotic gene conversion. Nature 338, 87–90 (1989).

    Article  CAS  Google Scholar 

  16. Liu, J., Wu, T.-C. & Lichten, M. The location and structure of double-strand DMA breaks induced during yeast meiosis: evidence for a covalently linked DNA-protein intermediate. EMBO J. 14, 4599–4608 (1995).

    Article  CAS  Google Scholar 

  17. de Massy, B., Rocco, V. & Nicolas, A. The nucleotide mapping of DNA double-strand breaks at the CYS3 initiation site of meiotic recombination in Saccharomyces cerevisiae. EMBO J. 14, 4589–4598 (1995).

    Article  CAS  Google Scholar 

  18. Fan, Q., Xu, F. & Petes, T.D. Meiosis-specific double-strand DNA breaks at the HIS4 recombination hot spot in the yeast Saccharomyces cerevisiae: Control in cis and trans. Mol. Cell. Biol. 15, 1679–1688 (1995).

    Article  CAS  Google Scholar 

  19. Ohta, K., Shibata, T. & Nicolas, A. Changes in chromatin structure at recombination initiation sites during yeast meiosis. EMBO J. 13, 5754–5763 (1994).

    Article  CAS  Google Scholar 

  20. Wu, T.-C. & Lichten, M. Meiosis-induced double-strand break sites determined by yeast chromatin structure. Science 263, 515–518 (1994).

    Article  CAS  Google Scholar 

  21. Wu, T.-C. & Lichten, M. Factors that affect the location and frequency of meiosis-induced double-strand breaks in Saccharomyces cerevisiae. Genetics 140, 55–66 (1995).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Haber, J.E. Exploring the pathways of homologous recombination. Curr.Biol. 4, 401–412 (1992).

    Article  CAS  Google Scholar 

  23. Hugerat, Y, Spencer, F., Zenvirth, D. & Simchen, G. A versatile method for efficient YAC transfer between any two strains. Genomics 22, 108–117 (1994).

    Article  CAS  Google Scholar 

  24. Walker, A.P., Chelly, J. & Love, D.R. A YAC contig in Xp21 containing the adrenal hypoplasia congenita and glycerol kinase deficiency genes. Hum. Mol. Genet. 1, 579–585 (1992).

    Article  CAS  Google Scholar 

  25. Sherman, F. Getting started with yeast. Meth. Enzymol. 194, 3–21 (1991).

    Article  CAS  Google Scholar 

  26. Kassir, Y. & Simchen, G. Monitoring meiosis and sporulation in Saccharomyces cerevisiae. Meth. Enzymol. 194, 94–110 (1991).

    Article  CAS  Google Scholar 

  27. Cao, L., Alani, E. & Kleckner, N. A pathway for generation and processing of double-strand breaks during meiotic recombination in S. cerevisiae. Cell 61, 1089–1101 (1990).

    Article  CAS  Google Scholar 

  28. de Massy, B. Baudat, F. & Nicolas, A. Initiation of recombination in Saccharomyces cerevisiae haploid meiosis. Proc. Natl. Acad. Sci. USA 91, 11929–11933 (1994).

    Article  CAS  Google Scholar 

  29. Alani, E., Padmore, R. & Kleckner, N. Analysis of wild-type and rad50 mutants of yeast suggests an intimate relationship between meiotic chromosome synapsis and recombination. Cell 61, 419–436 (1990).

    Article  CAS  Google Scholar 

  30. Sun, H., Treco, D. & Szostak, J.W. Extensive 3′-overhanging, single-stranded DNA associated with the meiosis-specific double-strand breaks at the ARG4 recombination initiation site. Cell 64, 1155–1161 (1991).

    Article  CAS  Google Scholar 

  31. 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 236, 806–612 (1987).

    Article  CAS  Google Scholar 

  32. Bussey, H. et al. The nucleotide sequence of chromosome I from Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 92, 3809–3813 (1995)

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Amir Sherman

    Present address: Whitehead Institute, Nine Cambridge Center, Cambridge, Massachusetts, 02142, USA

  2. Shoshana Klein and Drora Zenvirth: S.K. & D.Z. contributed equally to the work.

Authors and Affiliations

  1. Department of Genetics, Institute of Life Sciences, The Hebrew Universityof Jerusalem, Jerusalem, 91904, Israel

    Shoshana Klein, Drora Zenvirth, Amir Sherman & Giora Simchen

  2. Institute of Human Genetics, University ofHeidelberg, Heidelberg, Germany

    Karin Ried & Gudrun Rappold

Authors
  1. Shoshana Klein
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Drora Zenvirth
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Amir Sherman
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Karin Ried
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Gudrun Rappold
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Giora Simchen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Giora Simchen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Klein, S., Zenvirth, D., Sherman, A. et al. Double–strand breaks on YACs during yeast meiosis may reflect meiotic recombination in the human genome. Nat Genet 13, 481–484 (1996). https://doi.org/10.1038/ng0896-481

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/ng0896-481

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing