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Molecular and Cellular Biology, May 2008, p. 2884-2895, Vol. 28, No. 9
0270-7306/08/$08.00+0     doi:10.1128/MCB.00949-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Changes in the Distributions and Dynamics of Polycomb Repressive Complexes during Embryonic Stem Cell Differentiation{triangledown} ,{dagger}

Xiaojun Ren, Claudius Vincenz, and Tom K. Kerppola*

Howard Hughes Medical Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, Michigan 48109-0650

Received 29 May 2007/ Returned for modification 5 September 2007/ Accepted 20 February 2008

Polycomb group (PcG) transcription regulatory proteins maintain cell identity by sustained repression of numerous genes. The differentiation of embryonic stem (ES) cells induces a genome-wide shift in PcG target gene expression. We investigated the effects of differentiation and protein interactions on CBX family PcG protein localization and dynamics by using fluorescence imaging. In mouse ES cells, different CBX proteins exhibited distinct distributions and mobilities. Most CBX proteins were enriched in foci known as Polycomb bodies. Focus formation did not affect CBX protein mobilities, and the foci dispersed during ES cell differentiation. The mobilities of CBX proteins increased upon the induction of differentiation and decreased as differentiation progressed. The deletion of the chromobox, which mediates interactions with RING1B, prevented the immobilization of CBX proteins. In contrast, the deletion of the chromodomain, which can bind trimethylated lysine 27 of histone H3, had little effect on CBX protein dynamics. The distributions and mobilities of most CBX proteins corresponded to those of CBX-RING1B complexes detected by using bimolecular fluorescence complementation analysis. Epigenetic reprogramming during ES cell differentiation is therefore associated with global changes in the subnuclear distributions and dynamics of CBX protein complexes.

* Corresponding author. Mailing address: Howard Hughes Medical Institute and Department of Biological Chemistry, University of Michigan, Ann Arbor, MI 48109-0650. Phone: (734) 764-3553. Fax: (734) 615-3397. E-mail: kerppola{at}umich.edu

{triangledown} Published ahead of print on 3 March 2008.

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

Molecular and Cellular Biology, May 2008, p. 2884-2895, Vol. 28, No. 9
0270-7306/08/$08.00+0     doi:10.1128/MCB.00949-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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