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Chromatin Immunoprecipitation-Polymerase Chain Reaction (ChIP-PCR) Detects Methylation, Acetylation, and Ubiquitylation in S. pombe

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Book cover Schizosaccharomyces pombe

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1721))

Abstract

The distribution of modified histones within the fission yeast Schizosaccharomyces pombe genome is ultimately dependent upon the transcriptional activity and in turn influences the ability of the polymerases to bind and progress through the chromatin template. The Chromatin Immunoprecipitation-Polymerase Chain Reaction (ChIP-PCR) method currently provides the highest resolution, accuracy, and reproducibility to characterize histones modifications within a defined region of the genome. The following protocol details the method applied to S. pombe.

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References

  1. Zentner GE, Henikoff S (2013) Regulation of nucleosome dynamics by histone modifications. Nat Struct Mol Biol 20:259–266

    Article  CAS  PubMed  Google Scholar 

  2. Conaway JW (2012) Introduction to theme “Chromatin, epigenetics, and transcription”. Annu Rev Biochem 81:61–64

    Article  CAS  PubMed  Google Scholar 

  3. Kato H, Goto DB, Martienssen RA, Urano T, Furukawa K, Murakami Y (2005) RNA polymerase II is required for RNAi-dependent heterochromatin assembly. Science 309:467–469

    Article  CAS  PubMed  Google Scholar 

  4. Woolcock KJ, Buhler M (2013) Nuclear organisation and RNAi in fission yeast. Curr Opin Cell Biol 25(3):372–377

    Article  CAS  PubMed  Google Scholar 

  5. Fantes PA, Hoffman CS (2016) A brief history of schizosaccharomyces pombe research: a perspective over the past 70 years. Genetics 203:621–629

    Article  PubMed  PubMed Central  Google Scholar 

  6. Hoffman CS, Wood V, Fantes PA (2015) An ancient yeast for young geneticists: a primer on the schizosaccharomyces pombe model system. Genetics 201:403–423

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Allshire RC, Nimmo ER, Ekwall K, Javerzat JP, Cranston G (1995) Mutations derepressing silent centromeric domains in fission yeast disrupt chromosome segregation. Genes Dev 9:218–233

    Article  CAS  PubMed  Google Scholar 

  8. Cam HP, Sugiyama T, Chen ES, Chen X, FitzGerald PC, Grewal SI (2005) Comprehensive analysis of heterochromatin- and RNAi-mediated epigenetic control of the fission yeast genome. Nat Genet 37:809–819

    Article  CAS  PubMed  Google Scholar 

  9. Ren B, Robert F, Wyrick JJ, Aparicio O, Jennings EG, Simon I, Zeitlinger J, Schreiber J, Hannett N, Kanin E, Volkert TL, Wilson CJ, Bell SP, Young RA (2000) Genome-wide location and function of DNA binding proteins. Science 290:2306–2309

    Article  CAS  PubMed  Google Scholar 

  10. Gilmour DS, Lis JT (1984) Detecting protein-DNA interactions in vivo: distribution of RNA polymerase on specific bacterial genes. Proc Natl Acad Sci U S A 81:4275–4279

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Lenglez S, Hermand D, Decottignies A (2010) Genome-wide mapping of nuclear mitochondrial DNA sequences links DNA replication origins to chromosomal double-strand break formation in Schizosaccharomyces pombe. Genome Res 20:1250–1261

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Materne P, Anandhakumar J, Migeot V, Soriano I, Yague-Sanz C, Hidalgo E, Mignion C, Quintales L, Antequera F, Hermand D (2015) Promoter nucleosome dynamics regulated by signaling through the CTD code. Elife 4:e09008

    Article  PubMed  PubMed Central  Google Scholar 

  13. Materne P, Vazquez E, Sanchez M, Yague-Sanz C, Anandhakumar J, Migeot V, Antequera F, Hermand D (2016) Histone H2B ubiquitylation represses gametogenesis by opposing RSC-dependent chromatin remodeling at the ste11 master regulator locus. Elife 5:e13500

    Article  PubMed  PubMed Central  Google Scholar 

  14. Maundrell K (1990) nmt1 of fission yeast. A highly transcribed gene completely repressed by thiamine. J Biol Chem 265:10857–10864

    CAS  PubMed  Google Scholar 

  15. Matsuo Y, Nishino K, Mizuno K, Akihiro T, Toda T, Matsuo Y, Kaino T, Kawamukai M (2013) Polypeptone induces dramatic cell lysis in ura4 deletion mutants of fission yeast. PLoS One 8:e59887

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. URPHYM-GEMO, The University of Namur, Namur, Belgium

    Valerie Migeot & Damien Hermand

Authors
  1. Valerie Migeot
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  2. Damien Hermand
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Corresponding author

Correspondence to Damien Hermand .

Editor information

Editors and Affiliations

  1. Department of Biological Sciences, Winston-Salem State University, Winston-Salem, North Carolina, USA

    Teresa L. Singleton

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Migeot, V., Hermand, D. (2018). Chromatin Immunoprecipitation-Polymerase Chain Reaction (ChIP-PCR) Detects Methylation, Acetylation, and Ubiquitylation in S. pombe . In: Singleton, T. (eds) Schizosaccharomyces pombe. Methods in Molecular Biology, vol 1721. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7546-4_3

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  • DOI: https://doi.org/10.1007/978-1-4939-7546-4_3

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7545-7

  • Online ISBN: 978-1-4939-7546-4

  • eBook Packages: Springer Protocols

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