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Meiosis pp 37–51Cite as

Assaying Chromosome Pairing by FISH Analysis of Spread Saccharomyces cerevisiae Nuclei

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 558))

Abstract

Fluorescent in situ hybridization (FISH) provides a powerful tool to study the localization of DNA sequences in relationship to one another. FISH has the advantage over other methods, notably use of GFP-tagged repressor/operator arrays, that an almost unlimited number of probes can be utilized without having to make new strains for each new locus one wants to study. Also, the number of sites that can be visualized at the same time is limited only by the number of fluorophores that are available and can be distinguished by the available microscope. Described here is a method for FISH analysis and its application to analysis of chromosome pairing during meiosis in S. cerevisiae.

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References

  1. Weiner, B.M., and Kleckner, N. (1994) Chromosome pair via multiple interstitial interactions before and during meiosis in yeast. Cell 77, 977–991.

    Article  PubMed  CAS  Google Scholar 

  2. Burgess, S.M., Kleckner, N., and Weiner, B.M. (1999) Somatic pairing of homologs in budding yeast: existence and modulation.Genes Dev. 13, 1627–1641.

    Article  PubMed  CAS  Google Scholar 

  3. Cha, R.S., Weiner, B.M., Keeney, S., Dekker, J., and Kleckner, N. (2000) Progression of meiotic DNA replication is modulated by interchromosomal interaction proteins, negatively by Spo11p and positively by Rec8p. Genes Dev. 14, 493–503.

    PubMed  CAS  Google Scholar 

  4. Tessé, S., Storlazzi, A., Kleckner, N., Gargano, S., and Zickler D. (2003) Localization and roles of Ski8p protein in Sordaria meiosis and delineation of three mechanistically distinct steps of meiotic homolog juxtaposition. Proc. Natl. Acad. Sci. USA 100, 12865–12870.

    Article  PubMed  Google Scholar 

  5. Trelles-Sticken, E., Dresser, M.E., and Scherthan, H. (2000) Meiotic telomere protein Ndj1p is required for meiosis-specific telomere distribution, bouquet formation and efficient homologue pairing. J. Cell Biol. 2, 95–106.

    Article  Google Scholar 

  6. Padmore, R., Cao, L., and Kleckner, N. (1991) Temporal comparison of recombination and synaptonemal complex formation during meiosis in S. cerevisiae. Cell 66, 1239–1256.

    Article  PubMed  CAS  Google Scholar 

  7. Dresser, M.E., and Giroux, C.N. (1988) Meiotic behavior in spread preparations of yeast. J. Cell Biol. 106, 567–573.

    Article  PubMed  CAS  Google Scholar 

  8. Lorenz, A., Fuchs, J., Burger, R., and Loidl, J. (2003) Chromosome pairing does not contribute to nuclear architecture in vegetative yeast cells. Eukaryot. Cell 2, 856–866.

    Article  PubMed  CAS  Google Scholar 

  9. Singer, R.H., Lawrence, J.B., and Villnave, C. (1986) Optimization of in situ hybridization using isotopic and nonisotopic detection methods.BioTechniques 4, 230–250.

    CAS  Google Scholar 

  10. Jin, Q., Trelles-Sticken, E., Scherthan, H., and Loidl, J. (1998) Yeast nuclei display prominent centromere clustering that is reduced in nondividing cells and in meiotic prophase. J. Cell Biol. 141, 21–29.

    Article  PubMed  CAS  Google Scholar 

  11. Burgess, S.M., and Kleckner, N. (1999) Collisions between yeast chromosomal loci in vivo are governed by three layers of organization. Genes Dev. 13, 1871–1883.

    Article  PubMed  CAS  Google Scholar 

  12. Neale, M.J., Ramachandran, M., Trelles-Sticken, E., Scherthan, H., and Goldman, A.S. (2002) Wild-type levels of Spo11-induced DSBs are required for normal single-strand resection during meiosis. Mol. Cell 9, 835–846.

    Article  PubMed  CAS  Google Scholar 

  13. Guacci, V., Hogan, E., and Koshland, D. (1994) Chromosome condensation and sister chromatid pairing in budding yeast. J. Cell Biol. 125, 517–530.

    Article  PubMed  CAS  Google Scholar 

  14. Klein, F., Mahr, P., Galova, M., Buonomo, S.B., Michaelis, C., Nairz, K., and Nasmyth, K. (1999) A central role for cohesins in sister chromatid cohesion, formation of axial elements, and recombination during yeast meiosis. Cell 98, 91–103.

    Article  PubMed  CAS  Google Scholar 

  15. Bailis, J.M., and Roeder, G.S. (1998) Synaptonemal complex morphogenesis and sister-chromatid cohesion require Mek1-dependent phosphorylation of a meiotic chromosomal protein. Genes Dev. 12, 3551–3563.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

The above methods were developed in the laboratory of N. Kleckner, supported by NIH grants RO1-GM044794 and GM025326. We thank Neil Hunter and Ruth Padmore for particularly important contributions.

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Weiner, B.M., Kleckner, N. (2009). Assaying Chromosome Pairing by FISH Analysis of Spread Saccharomyces cerevisiae Nuclei. In: Keeney, S. (eds) Meiosis. Methods in Molecular Biology, vol 558. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-103-5_3

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  • DOI: https://doi.org/10.1007/978-1-60761-103-5_3

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-60761-102-8

  • Online ISBN: 978-1-60761-103-5

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