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Truncated HP1 lacking a functional chromodomain induces heterochromatinization upon in vivo targeting

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Abstract

Packaging of the eukaryotic genome into higher order chromatin structures is tightly related to gene expression. Pericentromeric heterochromatin is typified by accumulations of heterochromatin protein 1 (HP1), methylation of histone H3 at lysine 9 (MeH3K9) and global histone deacetylation. HP1 interacts with chromatin by binding to MeH3K9 through the chromodomain (CD). HP1 dimerizes with itself and binds a variety of proteins through its chromoshadow domain. We have analyzed at the single cell level whether HP1 lacking its functional CD is able to induce heterochromatinization in vivo. We used a lac-operator array-based system in mammalian cells to target EGFP-lac repressor tagged truncated HP1α and HP1β to a lac operator containing gene-amplified chromosome region in living cells. After targeting truncated HP1α or HP1β we observe enhanced tri-MeH3K9 and recruitment of endogenous HP1α and HP1β to the chromosome region. We show that CD-less HP1α can induce chromatin condensation, whereas the effect of truncated HP1β is less pronounced. Our results demonstrate that after lac repressor-mediated targeting, HP1α and HP1β without a functional CD are able to induce heterochromatinization.

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Acknowledgements

We are grateful to Dr. P.B. Singh, for kindly providing us with antibodies used in the present study. Confocal microscopy was performed at the Centre for Advanced Microscopy, we gratefully thank Dr. E.M.M. Manders for expert assistance. This work was supported by an ALW-NWO PULS and VIDI grant to PJV (project numbers PULS/33-98l/805-48011 and VIDI2003/03921/ALW/016.041.311).

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Authors and Affiliations

  1. Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, University of Amsterdam, Kruislaan 318, 1098 SM , Amsterdam, The Netherlands

    Maartje C. Brink, Yme van der Velden, Julio Mateos-Langerak, Roel van Driel & Pernette J. Verschure

  2. Center for Mathematics and Computer Science, CWI, Kruislaan 413, 1098, SJ, Amsterdam, The Netherlands

    Wim de Leeuw

  3. Department of Cell and Structural Biology, University of Illinois, Urbana-Champaign, Urbana, IL, 61801, USA

    Andrew S. Belmont

Authors
  1. Maartje C. Brink
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  2. Yme van der Velden
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  3. Wim de Leeuw
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  4. Julio Mateos-Langerak
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  5. Andrew S. Belmont
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  6. Roel van Driel
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  7. Pernette J. Verschure
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Correspondence to Pernette J. Verschure.

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Brink, M.C., van der Velden, Y., de Leeuw, W. et al. Truncated HP1 lacking a functional chromodomain induces heterochromatinization upon in vivo targeting. Histochem Cell Biol 125, 53–61 (2006). https://doi.org/10.1007/s00418-005-0088-7

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  • DOI: https://doi.org/10.1007/s00418-005-0088-7

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