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PMID: 19462008 Published · epublish English Journal Article Research Support, Non-U.S. Gov't

Polycomb mediated epigenetic silencing and replication timing at the INK4a/ARF locus during senescence.

PloS one ·Vol. 4 ·No. 5 ·2009-05-20 ·Pages e5622

Agherbi H, Gaussmann-Wenger A, Verthuy C, Chasson L, Serrano M, Djabali M

Abstract

The INK4/ARF locus encodes three tumor suppressor genes (p15(Ink4b), Arf and p16(Ink4a)) and is frequently inactivated in a large number of human cancers. Mechanisms regulating INK4/ARF expression are not fully characterized. Here we show that in young proliferating embryonic fibroblasts (MEFs) the Polycomb Repressive Complex 2 (PRC2) member EZH2 together with PRC1 members BMI1 and M33 are strongly expressed and localized at the INK4/ARF regulatory domain (RD) identified as a DNA replication origin. When cells enter senescence the binding to RD of both PRC1 and PRC2 complexes is lost leading to a decreased level of histone H3K27 trimethylation (H3K27me3). This loss is accompanied with an increased expression of the histone demethylase Jmjd3 and with the recruitment of the MLL1 protein, and correlates with the expression of the Ink4a/Arf genes. Moreover, we show that the Polycomb protein BMI1 interacts with CDC6, an essential regulator of DNA replication in eukaryotic cells. Finally, we demonstrate that Polycomb proteins and associated epigenetic marks are crucial for the control of the replication timing of the INK4a/ARF locus during senescence. We identified the replication licencing factor CDC6 as a new partner of the Polycomb group member BMI1. Our results suggest that in young cells Polycomb proteins are recruited to the INK4/ARF locus through CDC6 and the resulting silent locus is replicated during late S-phase. Upon senescence, Jmjd3 is overexpressed and the MLL1 protein is recruited to the locus provoking the dissociation of Polycomb from the INK4/ARF locus, its transcriptional activation and its replication during early S-phase. Together, these results provide a unified model that integrates replication, transcription and epigenetics at the INK4/ARF locus.

MeSH Terms
Animals Cell Cycle Proteins/metabolism Cellular Senescence Cyclin-Dependent Kinase Inhibitor p16/chemistry,genetics,metabolism DNA Replication Timing Embryo, Mammalian/cytology Enhancer of Zeste Homolog 2 Protein Fibroblasts/cytology,metabolism Gene Silencing Histone-Lysine N-Methyltransferase/metabolism Histones/metabolism Methylation Mice Models, Biological Myeloid-Lymphoid Leukemia Protein/metabolism NIH 3T3 Cells Nuclear Proteins/metabolism Polycomb Repressive Complex 1 Polycomb Repressive Complex 2 Polycomb-Group Proteins Protein Binding Protein Structure, Tertiary Proto-Oncogene Proteins/metabolism Repressor Proteins/metabolism
Chemicals
Bmi1 protein, mouse CDC6 protein, mouse Cbx2 protein, mouse Cell Cycle Proteins Cyclin-Dependent Kinase Inhibitor p16 Histones Nuclear Proteins Polycomb-Group Proteins Proto-Oncogene Proteins Repressor Proteins Myeloid-Lymphoid Leukemia Protein Enhancer of Zeste Homolog 2 Protein Ezh2 protein, mouse Histone-Lysine N-Methyltransferase Polycomb Repressive Complex 2 Polycomb Repressive Complex 1
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Agherbi Hanane
Centre d'Immunologie INSERM-CNRS de Marseille Luminy, Marseille, France.
Gaussmann-Wenger Anne
Verthuy Christophe
Chasson Lionel
Serrano Manuel
Djabali Malek
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Article Info
Journal
PloS one
Abbr.
PLoS One
ISSN
1932-6203
DOI
10.1371/journal.pone.0005622
Published
2009-05-20
Epub
2009-00-20
Pages
e5622
Language
English
Region
United States
NLM ID
101285081
PMCID
PMC2680618
Subset
IM
External Links
PubMed source DOI: 10.1371/journal.pone.0005622 PMC Full Text View journal details
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