Highly specific antibodies determine histone acetylation site usage in yeast heterochromatin and euchromatin.
We have developed a highly specific antibody set for acetylation sites in yeast histones H4 (K5, K8, K12, and K16); H3 (K9, K14, K18, K23, and K27); H2A (K7); and H2B (K11 and K16). Since ELISA does not assure antibody specificity in chromatin immunoprecipitation, we have employed additional screens against the respective histone mutations. We now show that telomeric and silent mating locus heterochromatin is hypoacetylated at all histone sites. At the INO1 promoter, RPD3 is required for strongly deacetylating all sites except H4 K16, ESA1 for acetylating H2A, H2B, and H4 sites except H4 K16, and GCN5 for acetylating H2B and H3 sites except H3 K14. These data uncover the in vivo usage of acetylation sites in heterochromatin and euchromatin.
Suka
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N
Department of Biological Chemistry, UCLA School of Medicine and, The Molecular Biology Institute, University of California, Los Angeles 90095, USA.
Suka
Y
Y
Carmen
A A
AA
Wu
J
J
Grunstein
M
M
D000107:Acetylation; D000123:Acetyltransferases / Q000378:metabolism; D000818:Animals; D000908:Antibodies, Fungal / Q000276:immunology / Q000378:metabolism*; D018797:Cell Cycle Proteins; D004797:Enzyme-Linked Immunosorbent Assay; D022041:Euchromatin / Q000276:immunology / Q000378:metabolism*; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D006570:Heterochromatin / Q000276:immunology / Q000378:metabolism*; D051548:Histone Acetyltransferases; D006657:Histones / Q000378:metabolism*; D047908:Intracellular Signaling Peptides and Proteins; D009154:Mutation; D011233:Precipitin Tests / Q000379:methods; D011401:Promoter Regions, Genetic / Q000235:genetics; D011494:Protein Kinases / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases; D029701:Saccharomyces cerevisiae Proteins; D015003:Yeasts / Q000235:genetics / Q000276:immunology / Q000378:metabolism*
D000908:Antibodies, Fungal; D018797:Cell Cycle Proteins; D022041:Euchromatin; D005656:Fungal Proteins; D006570:Heterochromatin; D006657:Histones; D047908:Intracellular Signaling Peptides and Proteins; D029701:Saccharomyces cerevisiae Proteins; D000123:Acetyltransferases; D051548:Histone Acetyltransferases; D011494:Protein Kinases; C064979:IME2 protein, S cerevisiae; D017346:Protein Serine-Threonine Kinases