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Molecular and Cellular Biology, December 2005, p. 11047-11058, Vol. 25, No. 24
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.24.11047-11058.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Association of BMI1 with Polycomb Bodies Is Dynamic and Requires PRC2/EZH2 and the Maintenance DNA Methyltransferase DNMT1§

Inmaculada Hernández-Muñoz ,1,{dagger},{ddagger} Panthea Taghavi,1,{dagger} Coenraad Kuijl,2 Jacques Neefjes,2 and Maarten van Lohuizen1*

Division of Molecular Genetics,1 Division of Tumor Biology, The Netherlands Cancer Institute, 1066 CX Amsterdam, The Netherlands2

Received 13 April 2005/ Returned for modification 10 June 2005/ Accepted 19 September 2005

Polycomb group (PcG) proteins are epigenetic chromatin modifiers involved in heritable gene repression. Two main PcG complexes have been characterized. Polycomb repressive complex 2 (PRC2) is thought to be involved in the initiation of gene silencing, whereas Polycomb repressive complex 1 (PRC1) is implicated in the stable maintenance of gene repression. Here, we investigate the kinetic properties of the binding of one of the PRC1 core components, BMI1, with PcG bodies. PcG bodies are unique nuclear structures located on regions of pericentric heterochromatin, found to be the site of accumulation of PcG complexes in different cell lines. We report the presence of at least two kinetically different pools of BMI1, a highly dynamic and a less dynamic fraction, which may reflect BMI1 pools with different binding capacities to these stable heterochromatin domains. Interestingly, PRC2 members EED and EZH2 appear to be essential for BMI1 recruitment to the PcG bodies. Furthermore, we demonstrate that the maintenance DNA methyltransferase DNMT1 is necessary for proper PcG body assembly independent of DNMT-associated histone deacetylase activity. Together, these results provide new insights in the mechanism for regulation of chromatin silencing by PcG proteins and suggest a highly regulated recruitment of PRC1 to chromatin.

* Corresponding author. Mailing address: Division of Molecular Genetics, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands. Phone: 31 20 512 2030. Fax: 31 20 512 2011. E-mail: m.v.lohuizen{at}nki.nl.

§ Supplemental material for this article may be found at http://mcb.asm.org./.

{dagger} I.H.-M. and P.T. contributed equally to this work.

{ddagger} Present address: Unidad de Biologia Celular y Molecular, Instituto Municipal de Investigaciones Medicas, 08003 Barcelona, Spain.

Molecular and Cellular Biology, December 2005, p. 11047-11058, Vol. 25, No. 24
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.24.11047-11058.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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