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First published online 10 May 2006
doi: 10.1242/dev.02405

Development 133, 2371-2381 (2006)
Published by The Company of Biologists 2006
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Distinct roles of Polycomb group gene products in transcriptionally repressed and active domains of Hoxb8

Yu-ichi Fujimura1,*, Kyo-ichi Isono1,*, Miguel Vidal2, Mitsuhiro Endoh1, Hiroshi Kajita1, Yoko Mizutani-Koseki1, Yoshihiro Takihara3, Maarten van Lohuizen4, Arie Otte5, Thomas Jenuwein6, Jacqueline Deschamps7 and Haruhiko Koseki1,{dagger}

1 RIKEN Research Center for Allergy and Immunology, 1-7-22 Suehiro, Tsurumi-ku, Yokohama 230-0045, Japan.
2 Centro de Investigaciones Biologicas, Department of Developmental and Cell Biology, Ramiro de Maeztu 9, 28040 Madrid, Spain.
3 Department of Stem Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima 734-8553, Japan.
4 Division of Molecular Genetics, The Netherlands Cancer Institute, 1066CX Amsterdam, The Netherlands.
5 Swammerdam Institute for Life Sciences, University of Amsterdam, Kruislaan 406, 1098 SM Amsterdam, The Netherlands.
6 Research Institute of Molecular Pathology, The Vienna Biocenter, Dr Bohrgasse 7, A-1030 Vienna, Austria.
7 Hubrecht Laboratory, Uppsalalaan 8 3584CT Utrecht, The Netherlands.

{dagger} Author for correspondence (e-mail: fujimara{at}rcai.riken.jp)

Accepted 12 April 2006

To address the molecular mechanisms underlying Polycomb group (PcG)-mediated repression of Hox gene expression, we have focused on the binding patterns of PcG gene products to the flanking regions of the Hoxb8 gene in expressing and non-expressing tissues. In parallel, we followed the distribution of histone marks of transcriptionally active H3 acetylated on lysine 9 (H3-K9) and methylated on lysine 4 (H3-K4), and of transcriptionally inactive chromatin trimethylated on lysine 27 (H3-K27). Chromatin immunoprecipitation revealed that the association of PcG proteins, and H3-K9 acetylation and H3-K27 trimethylation around Hoxb8 were distinct in tissues expressing and not expressing the gene. We show that developmental changes of these epigenetic marks temporally coincide with the misexpression of Hox genes in PcG mutants. Functional analyses, using mutant alleles impairing the PcG class 2 component Rnf2 or the Suz12 mutation decreasing H3-K27 trimethylation, revealed that interactions between class 1 and class 2 PcG complexes, mediated by trimethylated H3-K27, play decisive roles in the maintenance of Hox gene repression outside their expression domain. Within the expression domains, class 2 PcG complexes appeared to maintain the transcriptionally active status via profound regulation of H3-K9 acetylation. The present study indicates distinct roles for class 2 PcG complexes in transcriptionally repressed and active domains of Hoxb8 gene.

Key words: Polycomb, Hox, Mouse, Chromatin, Immunoprecipitation


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