Highly specific antibodies determine histone acetylation site usage in yeast heterochromatin and euchromatin
Highly specific antibodies determine histone acetylation site usage in yeast heterochromatin and euchromatin
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DOI：
10.1016/s1097-2765(01)00301-x
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发表时间：
2001-08-01
期刊：
影响因子：
16
通讯作者：
Grunstein, M
中科院分区：
文献类型：
--
作者：
Suka, N;Suka, Y;Grunstein, M
我们已经开发了针对酵母组蛋白H4（K5、K8、K12和K16）; H3（K9、K14、K18、K23和K27）; H2 A（K7）;和H213（K1 I和K16）中乙酰化位点的高度特异性抗体集。由于ELISA不能确保染色质免疫沉淀中的抗体特异性，我们已经采用了针对相应组蛋白突变的额外筛选。我们现在表明，端粒和沉默交配位点异染色质是hypoacetylated在所有组蛋白网站。在INO 1启动子处，RPD 3是使除H4 K16之外的所有位点强烈脱乙酰化所必需的，ESA 1是使除H4 K16之外的H2 A、H2 B和H4位点乙酰化所必需的，GCN 5是使除H3 K14之外的H213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的体内使用。
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 H213 (K1I 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 H213 and H3 sites except H3 K14. These data uncover the in vivo usage of acetylation sites in heterochromatin and euchromatin.