A Map of General and Specialized Chromatin Readers in Mouse Tissues Generated by Label-free Interaction Proteomics

Research output: Contribution to journalJournal articleResearchpeer-review

  • H.C. Eberl
  • M. Mann
  • C.G. Spruijt
  • M. Vermeulen
  • C.D. Kelstrup
Posttranslational modifications on core histones can serve as binding scaffolds for chromatin-associated proteins. Proteins that specifically bind to or "read" these modifications were previously identified in mass spectrometry-based proteomics screens based on stable isotope-labeling in cell lines. Here we describe a sensitive, label-free histone peptide pull-down technology with extracts of different mouse tissues. Applying this workflow to the classical activating and repressive epigenetic marks on histone H3, H3K4me3, and H3K9me3, we identified known and putative readers in extracts from brain, liver, kidney, and testis. A large class of proteins were specifically repelled by H3K4me3. Our screen reached near-saturation of direct interactors, most of which are ubiquitously expressed. In addition, it revealed a number of specialized readers in tissues such as testis. Apart from defining the chromatin interaction landscape in mouse tissues, our workflow can be used for peptides with different modifications and cell types of any organism.
Original languageEnglish
JournalMolecular Cell
Issue number2
Pages (from-to)368-378
Number of pages11
Publication statusPublished - 24 Jan 2013

ID: 47524117