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| Subject Categories:
Chromatin & Transcription
| Genome Stability & Dynamics
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The EMBO Journal
(2004) 23, 3516–3526, doi:10.1038/sj.emboj.7600362 Published online 12 August 2004
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| A CAF-1 dependent pool of HP1 during heterochromatin duplication |
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Jean-Pierre Quivy1, Danièle Roche1, Doris Kirschner1, 3, Hideaki Tagami2, 4, Yoshihiro Nakatani2 and Geneviève Almouzni1
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1 Institut Curie, Section de Recherche, UMR218 du CNRS, 26, Paris, France
2 Dana Farber Cancer Institute and Harvard Medical School, Boston, MA, USA
To whom correspondence should be addressed
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
3 Present address: Institut Pasteur, 25 rue du Dr. Roux, 75015 Paris, France
4 Present 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; Published online 12 August 2004.
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| Abstract |
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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. |
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| Keywords: chromatin assembly, heterochromatin, nuclear organization, replication in vivo |
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