Identification of a small molecule inhibitor of Sir2p
Identification of a small molecule inhibitor of Sir2p
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DOI：
10.1073/pnas.261574398
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发表时间：
2001-12-18
影响因子：
11.1
通讯作者：
Simon, JA
中科院分区：
文献类型：
--
作者：
Bedalov, A;Gatbonton, T;Simon, JA
Sir2p是酵母中染色质依赖性沉默所需的NAD（+）依赖性组蛋白去乙酰化酶。在Sir2p抑制剂的细胞筛选中，我们发现了一种化合物，splitomicin，它可以在酿酒酵母中产生sir2缺失突变体的条件表型。在药物存在下生长的细胞在端粒、沉默的配对型位点和核糖体DNA上存在沉默缺陷。此外，全基因组微阵列实验表明，splitomicin选择性抑制Sir2p。在体外实验中，splitomicin抑制Sir2蛋白的NAD+依赖性组蛋白去乙酰化酶活性（HIDA）。SIR2中赋予耐药性的突变指向该蛋白可能的乙酰化组蛋白尾部结合域。通过使用splitomicin作为化学遗传探针，我们证明Sir2p的连续HDA是维持非分裂细胞沉默状态所必需的。
Sir2p is an NAD(+)-dependent histone deacetylase required for chromatin-dependent silencing in yeast. In a cell-based screen for inhibitors of Sir2p, we identified a compound, splitomicin, that creates a conditional phenocopy of a sir2 deletion mutant in Saccharomyces cerevisiae. Cells grown in the presence of the drug have silencing defects at telomeres, silent mating-type loci, and the ribosomal DNA. In addition, whole genome microarray experiments show that splitomicin selectively inhibits Sir2p. In vitro, splitomicin inhibits NAD+-dependent histone deacetylase activity (HIDA) of the Sir2 protein. Mutations in SIR2 that confer resistance to the drug map to the likely acetylated histone tail binding domain of the protein. By using splitomicin as a chemical genetic probe, we demonstrate that continuous HDA of Sir2p is required for maintaining a silenced state in nondividing cells.