Global histone acetylation and deacetylation in yeast

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Abstract

Histone acetyltransferases and deacetylases can be targeted to promoters to activate or repress genes. For example, the histone acetyltransferase GCN5 is part of a yeast multiprotein complex that is recruited by the DNA-binding activator protein GCN4 (refs 1, 2,3). The histone deacetylase RPD3 complex is recruited to DNA by the repressor UME6 (refs 4, 5); similar mechanisms exist in other eukaryotes6. However, deletion of RPD3 also increases expression of the PHO5 gene7 that is repressed by nucleosomes8,9, and regulated by GCN5 (ref. 10) but not by UME6. We have determined whether acetylation and deacetylation are promoter specific at PHO5, by using antibodies against acetylated lysine residues and chromatin immunoprecipitation to examine the acetylation state of a 4.25-kilobase region surrounding the PHO5 gene. Here we show that this region is acetylated extensively by ESA1 and GCN5 and deacetylated by HDA1 and RPD3, and that widespread histone modification affects three separate chromosomal regions examined, which total 22 kb. Our data indicate that targeted modification occurs in a background of global acetylation and deacetylation that not only reduces basal transcription, but also allows a rapid return to the initial state of acetylation when targeting is removed.

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Figure 1: Global acetylation and deacetylation of a chromosomal region containing PHO5.
Figure 2: RPD3 is dominant to HDA1 in its repression of PHO5 .
Figure 3: HDA1 disruption slows return to repression of PHO5 from activating conditions.
Figure 4: Global deacetylation and acetylation in two regions of chromosome 10.

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Acknowledgements

We thank L. Pillus for the esa1ts strain. We are grateful to the members of the Grunstein laboratory for critical comments and discussions throughout this work and are thankful to Y. Suka and A. Carmen for antibodies against histone sites of acetylation. M.V. acknowledges pre-doctoral support from the Dottorato di Ricerca in Genetica e Biologia Molecolare—Università degli Studi di Roma “la Sapienza”. This work was supported by a Public Health Service grant from the National Institutes of Health.

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Correspondence to Michael Grunstein.

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