Histone acetyltransferase inhibitor anacardic acid causes changes in global gene expression during in vitro Plasmodium falciparum development
Histone acetyltransferase inhibitor anacardic acid causes changes in global gene expression during in vitro Plasmodium falciparum development
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DOI：
10.1128/ec.00063-08
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发表时间：
2008-07-01
期刊：
影响因子：
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通讯作者：
Cui, Liwang
中科院分区：
文献类型：
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作者：
Cui, Long;Miao, Jun;Cui, Liwang
为了更好地了解组蛋白赖氨酸乙酰化在恶性疟原虫转录中的作用，我们试图使用羽扇豆酸(AA)来减弱组蛋白乙酰转移酶(HAT)的活性。结果表明，AA对重组PfGCN5的HAT活性具有可逆性和非竞争性抑制作用。AA对寄生虫核提取液中PfGCN5的活性有一定的抑制作用，但不影响组蛋白脱乙酰酶的活性。AA对氯喹敏感株和抗性株的生长均有抑制作用，其50%抑制浓度约为30 mU M。用20 mU M的AA处理寄生虫12h，对寄生虫的生长和大体形态无明显影响，但在K9和K14引起组蛋白H3的低乙酰化，而在K5、K8、K12和K16不引起H4的低乙酰化，提示对PfGCN5 HAT有抑制作用。基因芯片分析表明，AA处理导致271个寄生虫基因在晚期滋养体中的表达发生了两倍或更大的变化(类似于5%)，其中207个基因下调。基因表达的聚类分析表明，AA主要下调活性基因，且该基因库与H3K9乙酰化富集库显著重叠。我们进一步通过染色质免疫沉淀和实时定量聚合酶链式反应证明，AA处理减少了一组下调基因的假定启动子附近的乙酰化。本研究提示AA的杀寄生虫作用至少部分与其对PfGCN5 HAT的抑制有关，从而导致寄生虫转录程序的紊乱。
To better understand the role of histone lysine acetylation in transcription in Plasmodium falciparum, we sought to attenuate histone acetyltransferase (HAT) activity using anacardic acid (AA). We showed that AA reversibly and noncompetitively inhibited the HAT activity of recombinant PfGCN5. To a lesser extent, AA inhibited the PfGCN5 activity in parasite nuclear extracts but did not affect histone deacetylase activity. AA blocked the growth of both chloroquine-sensitive and -resistant strains, with a 50% inhibitory concentration of similar to 30 mu M. Treatment of the parasites with 20 mu M of AA for 12 h had no obvious effect on parasite growth or gross morphology but induced hypoacetylation of histone H3 at K9 and K14, but not H4 at K5, K8, K12, and K16, suggesting inhibition of the PfGCN5 HAT. Microarray analysis showed that this AA treatment resulted in twofold or greater change in the expression of 271 (similar to 5%) parasite genes in late trophozoites, among which 207 genes were downregulated. Cluster analysis of gene expression indicated that AA mostly downregulated active genes, and this gene pool significantly overlapped with that enriched for H3K9 acetylation. We further demonstrated by chromatin immunoprecipitation and real-time PCR that AA treatment reduced acetylation near the putative promoters of a set of downregulated genes. This study suggests that the parasiticidal effect of AA is at least partially associated with its inhibition of PfGCN5 HAT, resulting in the disturbance of the transcription program in the parasites.