Suka N, Suka Y, Carmen A A, Wu J, Grunstein M
Department of Biological Chemistry, UCLA School of Medicine and, The Molecular Biology Institute, University of California, Los Angeles 90095, USA.
Mol Cell. 2001 Aug;8(2):473-9. doi: 10.1016/s1097-2765(01)00301-x.
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.
我们已针对酵母组蛋白H4（K5、K8、K12和K16）、H3（K9、K14、K18、K23和K27）、H2A（K7）以及H2B（K11和K16）的乙酰化位点开发了一套高度特异性的抗体。由于酶联免疫吸附测定（ELISA）无法确保染色质免疫沉淀中抗体的特异性，我们针对各自的组蛋白突变进行了额外的筛选。我们现在表明，端粒和沉默交配位点异染色质在所有组蛋白位点均低乙酰化。在INO1启动子处，RPD3是除H4 K16外所有位点强力去乙酰化所必需的，ESA1是除H4 K16外H2A、H2B和H4位点乙酰化所必需的，而GCN5是除H3 K14外H2B和H3位点乙酰化所必需的。这些数据揭示了异染色质和常染色质中乙酰化位点在体内的使用情况。
