Histone H3 serine 10 phosphorylation by Aurora B causes HP1 dissociation from heterochromatin

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Abstract

Histones are subject to numerous post-translational modifications1. Some of these ‘epigenetic’ marks recruit proteins that modulate chromatin structure. For example, heterochromatin protein 1 (HP1) binds to histone H3 when its lysine 9 residue has been tri-methylated by the methyltransferase Suv39h (refs 2–6). During mitosis, H3 is also phosphorylated by the kinase Aurora B 7. Although H3 phosphorylation is a hallmark of mitosis, its function remains mysterious. It has been proposed that histone phosphorylation controls the binding of proteins to chromatin8, but any such mechanisms are unknown. Here we show that antibodies against mitotic chromosomal antigens that are associated with human autoimmune diseases9 specifically recognize H3 molecules that are modified by both tri-methylation of lysine 9 and phosphorylation of serine 10 (H3K9me3S10ph). The generation of H3K9me3S10ph depends on Suv39h and Aurora B, and occurs at pericentric heterochromatin during mitosis in different eukaryotes. Most HP1 typically dissociates from chromosomes during mitosis10,11,12, but if phosphorylation of H3 serine 10 is inhibited, HP1 remains chromosome-bound throughout mitosis. H3 phosphorylation by Aurora B is therefore part of a ‘methyl/phos switch’ mechanism8 that displaces HP1 and perhaps other proteins from mitotic heterochromatin.

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Figure 1: Characterization of the MCA1 serum.
Figure 2: Identification of the MCA1 epitope.
Figure 3: HP1α remains associated with mitotic chromosomes if Aurora B is inhibited.
Figure 4: H3 Ser 10 phosphorylation is necessary and sufficient to prevent binding of HP1α to H3 peptides.

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Acknowledgements

We are grateful to K. Mechtler and M. Madalinski for peptide synthesis, to S. Opravil and T. Jenuwein for H3K9me3 antibodies and Suv39h-deficient MEFs, to A. Musacchio for purified Aurora B–INCENP complex, to H. Saya for Aurora A antibodies, and to W. Pollock for collecting MCA sera sent to Gribbles Pathology. T.H. acknowledges a fellowship from the Japanese Society for the Promotion of the Science (JSPS). Research in the laboratory of J.-M.P. is supported by Boehringer Ingelheim, Wiener Wirtschaftsfoerderungsfonds (WWFF), the Sixth Framework Programme of the European Union via the Integrated Project Mitocheck, the Austrian Science Fund and the European Molecular Biology Organization (EMBO). Author Contributions B.-H.T identified MCA sera. T.H. and J.-M.P. conceived and designed the experiments. T.H. and J.J.L. performed the experiments and analysed the data. T.H. and J.-M.P. wrote the paper.

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Correspondence to Jan-Michael Peters.

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Supplementary information

Supplementary Figure S1

Characterization of MCA2 and MCA6. (JPG 365 kb)

Supplementary Figure S2

The MCA1 epitope is conserved among species. (JPG 243 kb)

Supplementary Figure S3

H3 phosphopeptides do not compete with MCA1 reactivity. (JPG 233 kb)

Supplementary Figure S4

Localization of HP1α in interphase and at different stages of mitosis. (JPG 236 kb)

Supplementary Figure S5

Inhibition of Aurora-B also affects association of HP1β and HP1γ with mitotic chromosomes. (JPG 268 kb)

Supplementary Figure S6

Depletion of Aurora-A and -B. 48 hours after the transfection of indicated siRNAs, cell extracts were analyzed by immunoblotting for Aurora-A and Aurora-B. (JPG 164 kb)

Supplementary Table

Characterization of all six MCA sera. (JPG 272 kb)

Supplementary Figure Legends

Text to accompany the above Supplementary Figures. (DOC 31 kb)

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