Truncated HP1 lacking a functional chromodomain induces heterochromatinization upon in vivo targeting.
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 HP1alpha and HP1beta to a lac operator containing gene-amplified chromosome region in living cells. After targeting truncated HP1alpha or HP1beta we observe enhanced tri-MeH3K9 and recruitment of endogenous HP1alpha and HP1beta to the chromosome region. We show that CD-less HP1alpha can induce chromatin condensation, whereas the effect of truncated HP1beta is less pronounced. Our results demonstrate that after lac repressor-mediated targeting, HP1alpha and HP1beta without a functional CD are able to induce heterochromatinization.
Brink
Maartje C
MC
Swammerdam Institute for Life Sciences, BioCentrum Amsterdam, University of Amsterdam, Kruislaan 318, 1098 SM, Amsterdam, The Netherlands.
van der Velden
Yme
Y
de Leeuw
Wim
W
Mateos-Langerak
Julio
J
Belmont
Andrew S
AS
van Driel
Roel
R
Verschure
Pernette J
PJ
D000818:Animals; D016466:CHO Cells; D002843:Chromatin / Q000235:genetics / Q000378:metabolism* / Q000648:ultrastructure*; D000090266:Chromobox Protein Homolog 5; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*; D006224:Cricetinae; D005455:Fluorescent Antibody Technique; D006570:Heterochromatin / Q000235:genetics / Q000378:metabolism*; D006657:Histones / Q000378:metabolism; D006801:Humans; D007090:Image Interpretation, Computer-Assisted; D008239:Lysine / Q000378:metabolism; D008745:Methylation; D018613:Microscopy, Confocal; D010957:Plasmids / Q000235:genetics
C000715593:CBX1 protein, human; C000715594:CBX5 protein, human; D002843:Chromatin; D002868:Chromosomal Proteins, Non-Histone; D006570:Heterochromatin; D006657:Histones; D000090266:Chromobox Protein Homolog 5; D008239:Lysine