Phosphorylation of CBX2 controls its nucleosome-binding specificity

Takayuki Kawaguchi, Shinichi Machida, Hitoshi Kurumizaka, Hideaki Tagami, Jun Ichi Nakayama

    Research output: Contribution to journalArticle

    9 Citations (Scopus)

    Abstract

    Chromobox 2 (CBX2), a component of polycomb repressive complex 1 (PRC1), binds lysine 27-methylated histone H3 (H3K27me3) via its chromodomain (CD) and plays a critical role in repressing developmentally regulated genes. The phosphorylation of CBX2 has been described in several studies, but the biological implications of this modification remain largely elusive. Here, we show that CBX2's phosphorylation plays an important role in its nucleosome binding. CBX2 is stably phosphorylated in vivo, and domain analysis showed that residues in CBX2's serine-rich (SR) region are the predominant phosphorylation sites. The serine residues in an SR region followed by an acidicresidue (AR) cluster coincide with the consensus target of casein kinase II (CK2), and CK2 efficiently phosphorylated the SR region in vitro. A nucleosome pulldown assay revealed that CK2-phosphorylated CBX2 had a high specificity for H3K27me3-modified nucleosomes. An electrophoretic mobility-shift assay showed that CK2-mediated phosphorylation diminished CBX2's AT-hook-associated DNA-binding activity. Mutant CBX2 lacking the SR region or its neighboring AR cluster failed to repress the transcription of p21, a gene targeted by PRC1. These results suggest that CBX2's phosphorylation is critical for its transcriptional repression of target genes.

    Original languageEnglish
    Pages (from-to)343-355
    Number of pages13
    JournalJournal of Biochemistry
    Volume162
    Issue number5
    DOIs
    Publication statusPublished - 2017 Jan 1

    Keywords

    • CBX2
    • chromodomain
    • histone methylation
    • phosphorylation
    • polycomb

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology

    Fingerprint Dive into the research topics of 'Phosphorylation of CBX2 controls its nucleosome-binding specificity'. Together they form a unique fingerprint.

  • Cite this

    Kawaguchi, T., Machida, S., Kurumizaka, H., Tagami, H., & Nakayama, J. I. (2017). Phosphorylation of CBX2 controls its nucleosome-binding specificity. Journal of Biochemistry, 162(5), 343-355. https://doi.org/10.1093/jb/mvx040