Heterochromatin formation involves changes in histone modifications in multiple cell cycles.pdfVIP

Heterochromatin formation involves changes in histone modifications over multiple cell generations Yael Katan-Khaykovich and Kevin Struhl* Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA, USA Stable, epigenetic inactivation of gene expression by silen- cing complexes involves a specialized heterochromatin structure, but the kinetics and pathway by which euchro- matin is converted to the stable heterochromatin state are poorly understood. Induction of heterochromatin in Saccharomyces cerevisiae by expression of the silencing protein Sir3 results in rapid loss of histone acetylation, whereas removal of euchromatic histone methylation occurs gradually through several cell generations. Unexpectedly, Sir3 binding and the degree of transcrip- tional repression gradually increase for 3–5 cell genera- tions, even though the intracellular level of Sir3 remains constant. Strains lacking Sas2 histone acetylase or the histone methylases that modify lysines 4 (Set1) or 79 (Dot1) of H3 display accelerated Sir3 accumulation at HMR or its spreading away from the telomere, suggesting that these histone modifications exert distinct inhibitory effects on heterochromatin formation. These findings sug- gest an ordered pathway of heterochromatin assembly, consisting of an early phase, driven by active enzymatic removal of histone acetylation and resulting in incomplete transcriptional silencing, followed by a slower maturation phase, in which gradual loss of histone methylation en- hances Sir association and silencing. Thus, the transition between euchromatin and heterochromatin is gradual and requires multiple cell division cycles. The EMBO Journal (2005) 24, 2138–2149. doi:10.1038/ sj.emboj.7600692; Published online 26 May 2005 Subject Categories: chromatin transcription Keywords: epigenetic transition; heterochromatin; histone methylation; Sir proteins; transcriptional silencing Introduction Epigenetically inheritable patterns of gene