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Dynamic Interactome of PRC2-EZH1 Complex Using Tandem-Affinity Purification and Quantitative Mass Spectrometry

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Polycomb Group Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2655))

Abstract

The Polycomb repressive complex 2 (PRC2) is a well-characterized chromatin regulator of transcription programs acting through H3K27me3 deposition. In mammals, there are two main versions of PRC2 complexes: PRC2-EZH2, which is prevalent in cycling cells, and PRC2-EZH1 where EZH1 replaces EZH2 in post-mitotic tissues. Stoichiometry of PRC2 complex is dynamically modulated during cellular differentiation and various stress conditions. Therefore, unraveling unique architecture of PRC2 complexes under specific biological context through comprehensive and quantitative characterization could provide insight into the underlying mechanistic molecular mechanism in regulation of transcription process. In this chapter, we describe an efficient method which combines tandem-affinity purification (TAP) with label-free quantitative proteomics strategy for studying PRC2-EZH1 complex architecture alterations and identifying novel protein regulators in post-mitotic C2C12 skeletal muscle cells.

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Authors and Affiliations

  1. King Abdullah University of Science and Technology, Biological and Environmental Sciences and Engineering Division, KAUST Environmental Epigenetics Research Program, Thuwal, Kingdom of Saudi Arabia

    Peng Liu, Francesco Della Valle & Valerio Orlando

  2. King Abdullah University of Science and Technology, Core Labs, Thuwal, Kingdom of Saudi Arabia

    Huoming Zhang

Authors
  1. Peng Liu
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  2. Huoming Zhang
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  3. Francesco Della Valle
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  4. Valerio Orlando
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Corresponding authors

Correspondence to Peng Liu or Valerio Orlando .

Editor information

Editors and Affiliations

  1. Institute of Biomedical Technologies, National Research Council, Milan, Italy

    Chiara Lanzuolo

  2. Istituto Nazionale di Genetica Molecolare “Romeo ed Enrica Invernizzi”, INGM, Milan, Italy

    Federica Marasca

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Liu, P., Zhang, H., Della Valle, F., Orlando, V. (2023). Dynamic Interactome of PRC2-EZH1 Complex Using Tandem-Affinity Purification and Quantitative Mass Spectrometry. In: Lanzuolo, C., Marasca, F. (eds) Polycomb Group Proteins. Methods in Molecular Biology, vol 2655. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3143-0_9

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  • DOI: https://doi.org/10.1007/978-1-0716-3143-0_9

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3142-3

  • Online ISBN: 978-1-0716-3143-0

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