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*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

55 Scopus citations


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 Nterminus 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.

Original languageEnglish (US)
Pages (from-to)12498-12511
Number of pages14
Issue number20
StatePublished - Nov 10 2014
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2014 The Author(s). Published by Oxford University Press on behalf of Nucleic Acids Research.

ASJC Scopus subject areas

  • Genetics


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