Advertisement

Meiosis pp 285-304 | Cite as

Genome-Wide High-Resolution Chromatin Immunoprecipitation of Meiotic Chromosomal Proteins in Saccharomyces cerevisiae

  • Kazuto Kugou
  • Kunihiro Ohta
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 557)

Abstract

Cooperative actions of chromosomal proteins play critical roles in the dynamics, structural transition, segregation, and maintenance of meiotic chromosomes. A high-resolution genome-tiling array combined with a chromatin immunoprecipitation assay (ChIP-chip) is a powerful tool for uncovering the precise and dynamic distributions of various proteins along meiotic chromosomes. In this chapter, we describe a method to map the binding sites of meiotic chromosomal proteins such as Spo11, Mre11, and Rec8 using the high-resolution ChIP-chip technology. This system provides us with informative knowledge on the interplay of meiotic chromosomal proteins.

Key words

Recombination meiosis ChIP-chip genome-tiling array DNA–protein interaction 

Notes

Acknowledgments

We would like to thank all the laboratory members of Ohta laboratory and Shibata distinguished senior scientist laboratory for helpful discussion. We also thank Dr. W. Lin for critically reading this manuscript and Ms. Y. Sakuma for technical assistance. This work was supported by grants from the following sources: basic research from the Bio-oriented Technology Research Advancement Institution (to T. Shibata and K. Ohta); grants-in-aid for scientific research on priority areas from the Ministry of Education, Science, Culture, & Sports, Japan (to K. Ohta).

References

  1. 1.
    Ren, B., Robert, F., Wyrick, J. J., Aparicio, O., Jennings, E. G., Simon, I., et al. (2000) Genome-Wide location and function of DNA binding proteins. Science 290, 2306–2309.PubMedCrossRefGoogle Scholar
  2. 2.
    Katou, Y., Kanoh, Y., Bando, M., Noguchi, H., Tanaka, H., Ashikari, T., et al. (2003) S-phase checkpoint proteins Tof1 and Mrc1 form a stable replication-pausing complex. Nature 424, 1078–1083.PubMedCrossRefGoogle Scholar
  3. 3.
    Lengronne, A., Katou, Y., Mori, S., Yokobayashi, S., Kelly, G. K., Ito, T., et al. (2004) Cohesin relocation from sites of chromosomal loading to places of convergent transcription. Nature 430, 573–578.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Kazuto Kugou
    • 1
  • Kunihiro Ohta
    • 1
  1. 1.Department of Life Sciences, Graduate School of Arts and SciencesThe University of Tokyo, and Shibata Distinguished Senior Scientist Laboratory, RIKEN Discovery Research InstituteJapan

Personalised recommendations