Duplication and Maintenance of Heterochromatin Domains

doi:10.1083/jcb.147.6.1153

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© The Rockefeller University Press, 0021-9525/1999//1153 $5.00
The Journal of Cell Biology, Volume 147, Number 6, , 1999 1153-1166

Original Article

Duplication and Maintenance of Heterochromatin Domains

Angela Taddei^a, Danièle Roche^a, Jean-Baptiste Sibarita^a, Bryan M. Turner^b, and Geneviève Almouzni^a

^a Institut Curie, Research section, UMR 144 et 218 du Centre National de la Recherche Scientifique (CNRS), 75248 Paris cedex 05, France
^b Anatomy Department, University of Birmingham Medical School, Edgbaston, Birmingham, B152TT United Kingdom
Institut Curie, Research section, UMR 144 et 218 du CNRS, 75248 Paris cedex 05, France.+33 (0)1 42 34 64 21+33 (0)1 42 34 64 10

almouzni{at}curie.fr

To investigate the mechanisms that assure the maintenance ofheterochromatin regions, we took advantage of the fact thatclusters of heterochromatin DNA replicate late in S phase andare processed in discrete foci with a characteristic nucleardistribution. At the light microscopy level, within these entities,we followed DNA synthesis, histone H4 acetylation, heterochromatinprotein 1 (Hp1 ${alpha}$ and -β), and chromatin assembly factor 1(CAF-1). During replication, Hp1 ${alpha}$ and -β domains of concentrationare stably maintained, whereas heterochromatin regions are enrichedin both CAF-1 and replication-specific acetylated isoforms ofhistone H4 (H4Ac 5 and 12). We defined a time window of 20 minfor the maintenance of this state. Furthermore, treatment withTrichostatin A (TSA), during and after replication, sustainsthe H4Ac 5 and 12 state in heterochromatin excluding H4Ac 8and 16. In comparison, early replication foci, at the same level,did not display any specific enrichment in H4Ac 5 and 12. Thesedata emphasize the specific importance for heterochromatin ofthe replication-associated H4 isoforms. We propose that perpetuationof heterochromatin involves self-maintenance factors, includinglocal concentration of Hp1 ${alpha}$ and -β, and that a degree ofplasticity is provided by the cycle of H4 acetylation/deacetylationassisted by CAF-1.

Key Words: heterochromatin • nuclear organization • histone H4 acetylation • chromatin assembly factor 1 • heterochromatin protein 1

. Abbreviations used in this paper: BiodU, Biotin-16-deoxyuridine;BrdU, 5-bromo-2'-deoxyuridine; CAF-1, chromatin assembly factor1; H4Ac n, histone H4 acetylated at lysine n; HAT, histone acetyltransferase; HDAC, histone deacetylase; HP1, heterochromatinprotein 1; pAb, polyclonal antibody; TSA, Trichostatin A.

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