Analysis of phosphorylation-dependent protein-protein interactions of histone H3

Rebecca Klingberg, Jan Oliver Jost, Michael Schümann, Kathy Ann Gelato, Wolfgang Fischle, Eberhard Krause, Dirk Schwarzer*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

14 Scopus citations

Abstract

Multiple posttranslational modifications (PTMs) of histone proteins including site-specific phosphorylation of serine and threonine residues govern the accessibility of chromatin. According to the histone code theory, PTMs recruit regulatory proteins or block their access to chromatin. Here, we report a general strategy for simultaneous analysis of both of these effects based on a SILAC MS scheme. We applied this approach for studying the biochemical role of phosphorylated S10 of histone H3. Differential pull-down experiments with H3-tails synthesized from l- and d-amino acids uncovered that histone acetyltransferase 1 (HAT1) and retinoblastoma-binding protein 7 (RBBP7) are part of the protein network, which interacts with the unmodified H3-tail. An additional H3-derived bait containing the nonhydrolyzable phospho-serine mimic phosphonomethylen-alanine (Pma) at S10 recruited several isoforms of the 14-3-3 family and blocked the recruitment of HAT1 and RBBP7 to the unmodified H3-tail. Our observations provide new insights into the many functions of H3S10 phosphorylation. In addition, the outlined methodology is generally applicable for studying specific binding partners of unmodified histone tails.

Original languageEnglish (US)
Pages (from-to)138-145
Number of pages8
JournalACS Chemical Biology
Volume10
Issue number1
DOIs
StatePublished - Jan 16 2015

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine

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