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The HP1–p150/CAF-1 interaction is required for pericentric heterochromatin replication and S-phase progression in mouse cells

Nature Structural & Molecular Biology volume 15, pages 972–979 (2008)Cite this article

Abstract

The heterochromatin protein 1 (HP1)-rich heterochromatin domains next to centromeres are crucial for chromosome segregation during mitosis. This mitotic function requires their faithful reproduction during the preceding S phase, a process whose mechanism and regulation are current puzzles. Here we show that p150, a subunit of chromatin assembly factor 1, has a key role in the replication of pericentric heterochromatin and S-phase progression in mouse cells, independently of its known function in histone deposition. By a combination of depletion and complementation assays in vivo, we link this unique function of p150 to its ability to interact with HP1. Absence of this functional interaction triggers S-phase arrest at the time of replication of pericentromeric heterochromatin, without eliciting known DNA-based checkpoint pathways. Notably, in cells lacking the histone methylases Suv39h, in which pericentric domains do not show HP1 accumulation, p150 is dispensable for S-phase progression.

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Figure 1: Depletion of p150 but not p60 leads to accumulation of cells in early S phase.
Figure 2: Nucleosomal organization in p150- and p60-depleted cells.
Figure 3: Depletion of p150 does not affect the rate of DNA elongation but impairs centromeric DNA replication.
Figure 4: Progression from early to mid-late S phase depends on interaction between p150 and HP1.
Figure 5: S-phase progression is not perturbed in Suv39h dn MEF cells.

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Acknowledgements

We thank J. Haber for critical reading of the manuscript and members of our unit for discussions (D. Ray-Gallet, S. Polo, M.K. Ray and J. Chow); S. Huart for quantitative co-localization analyses; N. Murzina (Department of Biochemistry, University of Cambridge) and A. Verreault (IRIC, Université de Montréal, Canada) for the gift of mouse GFPp150 constructs; M. Moné for establishing 3T3Flp cell lines stably expressing GFPp150 proteins (Institute of Molecular Cell Biology, Faculty for Earth and Life Sciences, Vrije Universiteit, Amsterdam). A.G. received support from Délégation Générale pour l'Armement (DGA/DSP/STTC/DT/SH) and Association pour la Recherche sur la Cancer. A.J.L.C. received support from la Ligue Nationale Contre le Cancer. This work was supported by la Ligue Nationale Contre le Cancer (Equipe labellisée la Ligue), the Institut Curie PIC Programs ('Retinoblastome' and 'Replication, Instabilité chromosomique et cancer'), the European Commission Network of Excellence Epigenome (LSHG-CT-2004-503433), ACI-2004-Cancéropole IdF 'Breast cancer and epigenetics, ANR 'CenRNA' NT05-4_42267 and ANR 'FaRC', PCV06_142302.

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  1. Jean-Pierre Quivy and Annabelle Gérard: These authors contributed equally to this work.

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  1. Institut Curie, Centre de Recherche, UMR218 du CNRS, 26, rue d'Ulm, Paris, 75248, cedex 05, France

    Jean-Pierre Quivy, Annabelle Gérard, Adam J L Cook, Danièle Roche & Geneviève Almouzni

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  1. Jean-Pierre Quivy
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Contributions

J.-P.Q. and G.A. conceived and designed the experiments; J.-P.Q., A.G., A.J.L.C. and D.R. performed the experiments; J.-P.Q., A.G., A.J.L.C., D.R. and G.A. analyzed the data; J.-P.Q., A.J.L.C. and G.A. wrote the paper.

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Correspondence to Geneviève Almouzni.

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Quivy, JP., Gérard, A., Cook, A. et al. The HP1–p150/CAF-1 interaction is required for pericentric heterochromatin replication and S-phase progression in mouse cells. Nat Struct Mol Biol 15, 972–979 (2008). https://doi.org/10.1038/nsmb.1470

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