Knock down of GCN5 histone acetyltransferase by siRNA decreases ethanol-induced histone acetylation and affects differential expression of genes in human hepatoma cells.
Knock down of GCN5 histone acetyltransferase by siRNA decreases ethanol-induced histone acetylation and affects differential expression of genes in human hepatoma cells.
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DOI：
10.1016/j.alcohol.2010.12.003
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发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Shukla SD
中科院分区：
文献类型：
--
作者：
Choudhury M;Pandey RS;Clemens DL;Davis JW;Lim RW;Shukla SD
我们研究了组蛋白乙酰转移酶（HAT） Gcn5是否参与乙醇诱导的组蛋白H3赖氨酸9 （H3AcK9）的乙酰化，并对基因表达有任何影响。以转染乙醇代谢酶乙醇脱氢酶1 （VA 13）的人肝癌HepG2细胞为研究对象。通过siRNA沉默敲低Gcn5，降低了Gcn5的mRNA和蛋白水平、HAT活性，并减弱了乙醇诱导的VA13细胞中的H3AcK9。Illumina基因芯片分析显示，940个转录本受到GCN5沉默或乙醇的影响。沉默导致891个转录本的差异表达（≥1.5倍上调或下调）。其中，与对照相比，492个转录本上调，399个转录本下调。使用更严格的阈值（≥2.5倍），来自GCN5沉默样本的阵列数据显示57个基因差异表达（39个上调，18个下调）。同样，乙醇引起57个转录本的差异调控，变化≥1.5倍（35个基因上调，22个基因下调）。进一步分析表明，乙醇处理和GCN5沉默中共有的8个基因受到差异调控。其中，SLC44A2（一种假定的胆碱转运蛋白）被乙醇显著上调（3倍），而GCN5沉默使其下调（1.5倍）。定量RT-PCR图谱证实了阵列的发现。该报告首次证明(a) GCN5对多个基因的表达有差异影响，(b)乙醇诱导的组蛋白h3 -赖氨酸9乙酰化是通过GCN5介导的，(c) GCN5参与了乙醇诱导的胆碱转运体SLC44A2的表达。
We have investigated whether Gcn5, a histone acetyltransferase (HAT), is involved in ethanol induced acetylation of histone H3 at lysine 9 (H3AcK9) and has any effect on the gene expression. Human hepatoma HepG2 cells transfected with ethanol metabolizing enzyme alcohol dehydrogenase 1 (VA 13 cells) were used. Knockdown of Gcn5 by siRNA silencing decreased mRNA and protein levels of GCN5, HAT activity, and also attenuated ethanol induced H3AcK9 in VA13 cells. Illumina gene microarray analysis using total RNA showed 940 transcripts affected by GCN5 silencing or ethanol. Silencing caused differential expression of 891 transcripts (≥ 1.5 fold up- or down- regulated). Among these, 492 transcripts were up- and 399 were down- regulated compared to their respective controls. Using a more stringent threshold (≥ 2.5 fold) the array data from GCN5 silenced samples showed 57 genes differentially expressed (39 up-regulated and 18 down-regulated). Likewise, ethanol caused differential regulation of 57 transcripts with ≥ 1.5 fold change (35 gene up-regulated and 22 down-regulated). Further analysis showed that eight genes were differentially regulated that were common for both ethanol treatment and GCN5 silencing. Among these, SLC44A2 (a putative choline transporter) was strikingly up-regulated by ethanol (3 fold), and GCN5 silencing down regulated it (1.5 fold). The quantitative RT-PCR profile corroborated the array findings. This report demonstrates for the first time that (a) GCN5 differentially affects expression of multiple genes, (b) ethanol induced histone H3-lysine 9 acetylation is mediated via GCN5 and (c) that GCN5 is involved in ethanol induced expression of the putative choline transporter SLC44A2.
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影响因子：
16
作者：
Suka, N;Suka, Y;Grunstein, M
通讯作者：
Grunstein, M
影响因子：
--
作者：
HAJJAR, JJ;BAKER, ER;TOMICIC, TK
通讯作者：
TOMICIC, TK
DOI：
10.1073/pnas.97.21.11303
发表时间：
2000-10-10
影响因子：
11.1
作者：
Yamauchi, T;Yamauchi, J;Nakatani, Y
通讯作者：
Nakatani, Y
影响因子：
11.4
作者：
Zhang, WZ;Bone, JR;Roth, SY
通讯作者：
Roth, SY
影响因子：
13.5
作者：
Clemens, DL;Forman, A;Tuma, DJ
通讯作者：
Tuma, DJ