Steady-state Levels of Histone Acetylation in Saccharomyces cerevisiae

Abstract

The importance of control of the levels of histone acetylation for the control of gene expression in eukaryotic chromatin is being elucidated, and the yeast Saccharomyces cerevisiae has proven to be an important model system. The level of histone acetylation in yeast is the highest known. However, only acetylation of H4 has been quantified, and reports reveal loss of acetylation in histone preparations. A chaotropic guanidine-based method for histone isolation from intact wild-type cells or from a single-step nuclear preparation with butyrate preserves acetylation of all core histones. Histone H4 has an average of more than 2 acetylated lysines per molecule, distributed over 4 sites. Histones H2A, H3, and H2B have 0. 2, approximately 2, and >2 acetylated lysines per molecule, respectively, distributed across 2, 5, and 6 sites. Thus, yeast nucleosomes carry, on average, 13 acetylated lysines per octamer, i. e. just above the threshold of 10-12 deduced for transcriptionally activated chromatin of animals, plants, and algae. Following M(r) 100,000 ultrafiltration in 2.5% acetic acid, yeast histone H3 was purified to homogeneity by reversed-phase high pressure liquid chromatography. Other core histones were obtained at 80-95% purity.