Meiosis pp 157-174 | Cite as

Analysis of Meiotic Chromosome-Associated Protein Dynamics Using Conditional Expression in Budding Yeast

Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1471)

Abstract

The visualization of meiotic chromosomes and their associated protein structures in both wild-type and mutant cells adds valuable insight into the molecular pathways that underlie reproductive cell formation. Here we describe basic methodology for visualizing meiotic chromosomes in a long-standing model organism for investigating the molecular and cell biology of meiosis, the budding yeast, S. cerevisiae. This chapter furthermore highlights a variety of conditional expression regimes that can be used to understand the dynamics and/or developmental constraints of chromosomal protein structures; such dynamic aspects of the macromolecular structures that mediate meiotic chromosome biology are typically not obvious from standard protein visualization experiments.

Key words

Meiosis Chromatin Synaptonemal complex Conditional expression Yeast 

References

  1. 1.
    Page SL, Hawley RS (2004) The genetics and molecular biology of the synaptonemal complex. Annu Rev Cell Dev Biol 20:525CrossRefPubMedGoogle Scholar
  2. 2.
    Zickler D, Kleckner N (1999) Meiotic chromosomes: integrating structure and function. Annu Rev Genet 33:603CrossRefPubMedGoogle Scholar
  3. 3.
    MacQueen AJ, Unpublished dataGoogle Scholar
  4. 4.
    Voelkel-Meiman K, Moustafa SS, Lefrancois P, Villeneuve AM, MacQueen AJ (2012) Full-length synaptonemal complex grows continuously during meiotic prophase in budding yeast. PLoS Genet 8:e1002993CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Voelkel-Meiman K et al (2013) SUMO localizes to the central element of synaptonemal complex and is required for the full synapsis of meiotic chromosomes in budding yeast. PLoS Genet 9:e1003837CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    White EJ, Cowan C, Cande WZ, Kaback DB (2004) In vivo analysis of synaptonemal complex formation during yeast meiosis. Genetics 167:51CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Benjamin KR, Zhang C, Shokat KM, Herskowitz I (2003) Control of landmark events in meiosis by the CDK Cdc28 and the meiosis-specific kinase Ime2. Genes Dev 17:1524CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Jambhekar A, Amon A (2008) Control of meiosis by respiration. Curr Biol 18:969CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Rockmill B (2009) Chromosome spreading and immunofluorescence methods in Saccharomyes cerevisiae. Methods Mol Biol 558:3CrossRefPubMedGoogle Scholar
  10. 10.
    Longtine MS et al (1998) Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. Yeast 14:953CrossRefPubMedGoogle Scholar
  11. 11.
    Xu L, Ajimura M, Padmore R, Klein C, Kleckner N (1995) NDT80, a meiosis-specific gene required for exit from pachytene in Saccharomyces cerevisiae. Mol Cell Biol 15:6572CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Wakeling AE, Dukes M, Bowler J (1991) A potent specific pure antiestrogen with clinical potential. Cancer Res 51:3867PubMedGoogle Scholar
  13. 13.
    Rockmill B, Roeder GS (1998) Telomere-mediated chromosome pairing during meiosis in budding yeast. Genes Dev 12:2574CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Loidl J, Klein F, Engebrecht J (1998) Genetic and morphological approaches for the analysis of meiotic chromosomes in yeast. Methods Cell Biol 53:257CrossRefPubMedGoogle Scholar
  15. 15.
    Hollingsworth NM, Goetsch L, Byers B (1990) The HOP1 gene encodes a meiosis-specific component of yeast chromosomes. Cell 61:73CrossRefPubMedGoogle Scholar
  16. 16.
    Smith AV, Roeder GS (1997) The yeast Red1 protein localizes to the cores of meiotic chromosomes. J Cell Biol 136:957CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Molecular Biology and BiochemistryWesleyan UniversityMiddletownUSA
  2. 2.Howard Hughes Medical Institute and Department of Molecular and Cell BiologyUniversity of California-BerkeleyBerkeleyUSA

Personalised recommendations