N-terminal phosphorylation of HP1α increases its nucleosome-binding specificity

Gohei Nishibuchi, Shinichi Machida, Akihisa Osakabe, Hiromu Murakoshi, Kyoko Hiragami-Hamada, Reiko Nakagawa, Wolfgang Fischle, Yoshifumi Nishimura, Hitoshi Kurumizaka, Hideaki Tagami, Jun ichi Nakayama

    研究成果: Article

    20 引用 (Scopus)

    抄録

    Heterochromatin protein 1 (HP1) is an evolutionarily conserved chromosomal protein that binds to lysine 9-methylated histone H3 (H3K9me), a hallmark of heterochromatin. Although HP1 phosphorylation has been described in several organisms, the biological implications of this modification remain largely elusive. Here we show that HP1's phosphorylation has a critical effect on its nucleosome binding properties. By in vitro phosphorylation assays and conventional chromatography, we demonstrated that casein kinase II (CK2) is the kinase primarily responsible for phosphorylating the N-terminus of human HP1α. Pull-down assays using in vitro-reconstituted nucleosomes showed that unmodified HP1α bound H3K9-methylated and H3K9-unmethylated nucleosomes with comparable affinity, whereas CK2-phosphorylated HP1α showed a high specificity for H3K9me3-modified nucleosomes. Electrophoretic mobility shift assays showed that CK2-mediated phosphorylation diminished HP1α's intrinsic DNA binding, which contributed to its H3K9me-independent nucleosome binding. CK2-mediated phosphorylation had a similar effect on the nucleosome-binding specificity of fly HP1a and S. pombe Swi6. These results suggested that HP1 phosphorylation has an evolutionarily conserved role in HP1's recognition of H3K9me-marked nucleosomes.

    元の言語English
    ページ(範囲)12498-12511
    ページ数14
    ジャーナルNucleic Acids Research
    42
    発行部数20
    DOI
    出版物ステータスPublished - 2014 11 10

    Fingerprint

    Nucleosomes
    Phosphorylation
    Casein Kinase II
    Heterochromatin
    Schizosaccharomyces
    Electrophoretic Mobility Shift Assay
    heterochromatin-specific nonhistone chromosomal protein HP-1
    Diptera
    Histones
    Lysine
    Chromatography
    Phosphotransferases
    DNA

    ASJC Scopus subject areas

    • Medicine(all)

    これを引用

    Nishibuchi, G., Machida, S., Osakabe, A., Murakoshi, H., Hiragami-Hamada, K., Nakagawa, R., ... Nakayama, J. I. (2014). N-terminal phosphorylation of HP1α increases its nucleosome-binding specificity. Nucleic Acids Research, 42(20), 12498-12511. https://doi.org/10.1093/nar/gku995

    N-terminal phosphorylation of HP1α increases its nucleosome-binding specificity. / Nishibuchi, Gohei; Machida, Shinichi; Osakabe, Akihisa; Murakoshi, Hiromu; Hiragami-Hamada, Kyoko; Nakagawa, Reiko; Fischle, Wolfgang; Nishimura, Yoshifumi; Kurumizaka, Hitoshi; Tagami, Hideaki; Nakayama, Jun ichi.

    :: Nucleic Acids Research, 巻 42, 番号 20, 10.11.2014, p. 12498-12511.

    研究成果: Article

    Nishibuchi, G, Machida, S, Osakabe, A, Murakoshi, H, Hiragami-Hamada, K, Nakagawa, R, Fischle, W, Nishimura, Y, Kurumizaka, H, Tagami, H & Nakayama, JI 2014, 'N-terminal phosphorylation of HP1α increases its nucleosome-binding specificity', Nucleic Acids Research, 巻. 42, 番号 20, pp. 12498-12511. https://doi.org/10.1093/nar/gku995
    Nishibuchi G, Machida S, Osakabe A, Murakoshi H, Hiragami-Hamada K, Nakagawa R その他. N-terminal phosphorylation of HP1α increases its nucleosome-binding specificity. Nucleic Acids Research. 2014 11 10;42(20):12498-12511. https://doi.org/10.1093/nar/gku995
    Nishibuchi, Gohei ; Machida, Shinichi ; Osakabe, Akihisa ; Murakoshi, Hiromu ; Hiragami-Hamada, Kyoko ; Nakagawa, Reiko ; Fischle, Wolfgang ; Nishimura, Yoshifumi ; Kurumizaka, Hitoshi ; Tagami, Hideaki ; Nakayama, Jun ichi. / N-terminal phosphorylation of HP1α increases its nucleosome-binding specificity. :: Nucleic Acids Research. 2014 ; 巻 42, 番号 20. pp. 12498-12511.
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