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Molecular and Cellular Biology, June 2005, p. 4552-4564, Vol. 25, No. 11
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.11.4552-4564.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

In Vivo HP1 Targeting Causes Large-Scale Chromatin Condensation and Enhanced Histone Lysine Methylation{dagger}

Pernette J. Verschure,1* Ineke van der Kraan,1 Wim de Leeuw,2 Johan van der Vlag,3 Anne E. Carpenter,4 Andrew S. Belmont,4 and Roel van Driel1

Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, University of Amsterdam, Kruislaan 318, 1098 SM Amsterdam, The Netherlands,1 Center for Mathematics and Computer Science, CWI, 1090 GB Amsterdam, The Netherlands,2 Nephrology Research Lab, Nijmegen Center for Molecular Life Sciences, Division of Nephrology, University Medical Center, Nijmegen, The Netherlands,3 Department of Cell and Structural Biology, University of Illinois, Urbana-Champaign, Urbana, Illinois 618014

Received 5 December 2004/ Returned for modification 20 January 2005/ Accepted 22 February 2005

Changes in chromatin structure are a key aspect in the epigenetic regulation of gene expression. We have used a lac operator array system to visualize by light microscopy the effect of heterochromatin protein 1 (HP1) {alpha} (HP1{alpha}) and HP1ß on large-scale chromatin structure in living mammalian cells. The structure of HP1, containing a chromodomain, a chromoshadow domain, and a hinge domain, allows it to bind to a variety of proteins. In vivo targeting of an enhanced green fluorescent protein-tagged HP1-lac repressor fusion to a lac operator-containing, gene-amplified chromosome region causes local condensation of the higher-order chromatin structure, recruitment of the histone methyltransferase SETDB1, and enhanced trimethylation of histone H3 lysine 9. Polycomb group proteins of both the HPC/HPH and the EED/EZH2 complexes, which are involved in the heritable repression of gene activity, are not recruited to the amplified chromosome region by HP1{alpha} and HP1ß in vivo targeting. HP1{alpha} targeting causes the recruitment of endogenous HP1ß to the chromatin region and vice versa, indicating a direct interaction between the two HP1 homologous proteins. Our findings indicate that HP1{alpha} and HP1ß targeting is sufficient to induce heterochromatin formation.

* Corresponding author. Mailing address: Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, University of Amsterdam, P.O. Box 94062, 1090 GB Amsterdam, The Netherlands. Phone: 31.20.525.5151. Fax: 31.20.525.7924. E-mail: pj.verschure{at}science.uva.nl.

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

Molecular and Cellular Biology, June 2005, p. 4552-4564, Vol. 25, No. 11
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.11.4552-4564.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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