Dissecting the biological functions of Drosophila histone deacetylases by RNA interference and transcriptional profiling
Dissecting the biological functions of Drosophila histone deacetylases by RNA interference and transcriptional profiling
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DOI：
10.1074/jbc.m511945200
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发表时间：
2006-06-30
影响因子：
4.8
通讯作者：
Steinkuhler, Christian
中科院分区：
文献类型：
--
作者：
Foglietti, Cristiana;Filocamo, Gessica;Steinkuhler, Christian
锌依赖性组蛋白脱乙酰酶（HDAC）是首先被鉴定为转录抑制复合物的组分的水解酶家族，其中它们通过使核心组蛋白的N-末端延伸处的赖氨酸残基脱乙酰化而起作用，从而影响转录。为了更深入地了解单个HDAC家族成员的生物学功能，我们在果蝇S2细胞中使用了RNA干扰与微阵列分析相结合。果蝇HDAC 1（DHDAC 1）的沉默，而不是其他DHDAC家族成员，导致组蛋白乙酰化增加。DHDAC 1或DHDAC 3的沉默导致细胞生长抑制和共同和不同基因组的转录失调。沉默DHDAC 2导致微管蛋白乙酰化水平增加，但与基因表达失调无关。在DHDAC 4和DHDACX沉默后，没有观察到表型的生长和基因表达的显著失调。DHDAC 1缺失或S2细胞暴露于小分子HDAC抑制剂阿司他汀均导致G（2）阻滞，并与显著重叠的基因表达特征相关，其中涉及核碱基和脂质代谢、DNA复制、细胞周期调控和信号转导的基因过度表达。这些基因中的大量也显示在失去辅阻遏物SIN 3后被去调节（Pile，L.一、Spellman，P.T.，Katzenberger，R.J.，和Wassarman，D. A.（2003）J.Biol.Chem.278，37840-37848）。我们得出以下结论。1)DHDAC 1和DHDAC-3在基因表达调控中具有不同的功能。2)在测试的条件下，DHDAC 2、DHDAC-4和DHDAC X在S2细胞中没有可检测的转录功能。3)DHDAC 1的缺失在很大程度上再现了阿司他汀的抗增殖和转录作用。4)DHDAC 1的脱乙酰酶活性显著地有助于SIN 3的阻遏物功能。
Zinc-dependent histone deacetylases (HDACs) are a family of hydrolases first identified as components of transcriptional repressor complexes, where they act by deacetylating lysine residues at the N-terminal extensions of core histones, thereby affecting transcription. To get more insight into the biological functions of the individual HDAC family members, we have used RNA interference in combination with microarray analysis in Drosophila S2 cells. Silencing of Drosophila HDAC1 (DHDAC1), but not of the other DHDAC family members, leads to increased histone acetylation. Silencing of either DHDAC1 or DHDAC3 leads to cell growth inhibition and deregulated transcription of both common and distinct groups of genes. Silencing DHDAC2 leads to increased tubulin acetylation levels but was not associated with a deregulation of gene expression. No growth of phenotype and no significant deregulation of gene expression was observed upon silencing of DHDAC4 and DHDACX. Loss of DHDAC1 or exposure of S2 cells to the small molecule HDAC inhibitor trichostatin both lead to a G(2) arrest and were associated with significantly overlapping gene expression signatures in which genes involved in nucleobase and lipid metabolism, DNA replication, cell cycle regulation, and signal transduction were over-represented. A large number of these genes were shown to also be deregulated upon loss of the co-repressor SIN3 (Pile, L. A., Spellman, P. T., Katzenberger, R.J., and Wassarman, D. A. (2003) J. Biol. Chem. 278, 37840-37848). We conclude the following. 1) DHDAC1 and -3 have distinct functions in the control of gene expression. 2) Under the tested conditions, DHDAC2, -4, and X have no detectable transcriptional functions in S2 cells. 3) The anti-proliferative and transcriptional effects of trichostatin are largely recapitulated by the loss of DHDAC1. 4) The deacetylase activity of DHDAC1 significantly contributes to the repressor function of SIN3.