High-resolution genome-wide mapping of histone modifications.. Saccharomyces cerevisiae

The expression patterns of eukaryotic genomes are controlled by their chromatin structure, consisting of nucleosome subunits in which DNA of approximately 146 bp is wrapped around a core of 8 histone molecules. Post-translational histone modifications play an essential role in modifying chromatin structure. Here we apply a combination of SAGE and chromatin immunoprecipitation (ChIP) protocols to determine the distribution of hyperacetylated histones H3 and H4 in the Saccharomyces cerevisiae genome. We call this approach genome-wide mapping technique (GMAT). Using GMAT, we find that the highest acetylation levels are detected in the 5' end of a gene's coding region, but not in the promoter. Furthermore, we show that the histone acetyltransferase, GCN5p, regulates H3 acetylation in the promoter and 5' end of the coding regions. These findings indicate that GMAT should find valuable applications in mapping target sites of chromatin-modifying enzymes. Keywords: SAGE-ChIP Overall design: Using histones from wild-type S. cerevisiae as well as from yeast with a GCN5 deletion, we generated four GMAT libraries from the ChIP DNA with the antibodies against diacetylated (K9/K14) histone H3 and tetra-acetylated histone H4 tails.