Article
- The EMBO Journal (2004) 23, 3516 - 3526
- doi:10.1038/sj.emboj.7600362
Published online: 12 August 2004
Subject Categories:
A CAF-1 dependent pool of HP1 during heterochromatin duplication
Jean-Pierre Quivy1, Danièle Roche1, Doris Kirschner1,a, Hideaki Tagami2,b, Yoshihiro Nakatani2 and Geneviève Almouzni1
- Institut Curie, Section de Recherche, UMR218 du CNRS, 26, Paris, France
- Dana Farber Cancer Institute and Harvard Medical School, Boston, MA, USA
Correspondence to:
Geneviève Almouzni, Institut Curie, Section de Recherche, UMR218 du CNRS, 26, rue d'Ulm, 75248 Paris cedex 05, France. Tel.: + 33 1 4234 6701/6706; Fax: +33 1 4633 3016; E-mail: genevieve.almouzni@curie.fr
aPresent address: Institut Pasteur, 25 rue du Dr. Roux, 75015 Paris, France
bPresent address: Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa, Nagoya, Aichi 464-8602, Japan
Received 3 June 2004; Accepted 15 July 2004
Abstract
To investigate how the complex organization of heterochromatin is reproduced at each replication cycle, we examined the fate of HP1-rich pericentric domains in mouse cells. We find that replication occurs mainly at the surface of these domains where both PCNA and chromatin assembly factor 1 (CAF-1) are located. Pulse–chase experiments combined with high-resolution analysis and 3D modeling show that within 90 min newly replicated DNA become internalized inside the domain. Remarkably, during this time period, a specific subset of HP1 molecules (
and 
) coinciding with CAF-1 and replicative sites is resistant to RNase treatment. Furthermore, these replication-associated HP1 molecules are detected in Suv39 knockout cells, which otherwise lack stable HP1 staining at pericentric heterochromatin. This replicative pool of HP1 molecules disappears completely following p150CAF-1 siRNA treatment. We conclude that during replication, the interaction of HP1 with p150CAF-1 is essential to promote delivery of HP1 molecules to heterochromatic sites, where they are subsequently retained by further interactions with methylated H3-K9 and RNA.
Keywords:
- chromatin assembly,
- heterochromatin,
- nuclear organization,
- replication in vivo
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