Histone demethylase LSD1-mediated repression of GATA-2 is critical for erythroid differentiation.
Histone demethylase LSD1-mediated repression of GATA-2 is critical for erythroid differentiation.
复制标题
组蛋白去甲基化酶 LSD1 介导的 GATA-2 抑制对于红细胞分化至关重要。
DOI：
10.2147/dddt.s81911
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发表时间：
2015
期刊：
影响因子：
--
通讯作者：
Hu X
中科院分区：
文献类型：
--
作者：
Guo Y;Fu X;Jin Y;Sun J;Liu Y;Huo B;Li X;Hu X
转录因子GATA-2主要在造血干、祖细胞中表达，并与红系特异性转录因子GATA-1相互作用，调节造血细胞的增殖和分化。在造血细胞分化过程中，GATA-2呈现动态表达模式，受多种转录因子调控。小鼠红白血病(MEL)或小鼠胚胎干细胞与病毒颗粒一起在4μg/mLPolybrene®存在下培养24-48小时，再用1μg/mL嘌r霉素筛选2周，建立了稳定的LSD1基因敲除细胞系。采用实时聚合酶链式反应(PCR)定量染色质免疫沉淀法(CHIP)检测TAL1转录因子是否与GATA-2基因1S启动子结合，以及LSD1是否与TAL1在1S启动子上共定位。利用序列芯片技术证实LSD1在红系分化过程中对H3K4me2基因GATA-2基因的调控作用。Western印迹分析检测蛋白表达。用AlamarBlue®比色法检测细胞增殖情况，在光密度(OD)570 nm和OD 600 nm处检测吸光度。在本研究中，我们发现LSD1在红系分化过程中调节GATA-2的表达。LSD1基因敲除导致GATA-2表达增加，并抑制MEL和胚胎干细胞的分化。此外，我们还证明LSD1通过组蛋白去甲基化与GATA-2基因的1S启动子结合并抑制GATA-2的表达。我们的数据显示，LSD1通过GATA-2基因的表观遗传修饰介导了红系分化。
The transcription factor GATA-2 is predominantly expressed in hematopoietic stem and progenitor cells and counteracts the erythroid-specific transcription factor GATA-1, to modulate the proliferation and differentiation of hematopoietic cells. During hematopoietic cell differentiation, GATA-2 exhibits dynamic expression patterns, which are regulated by multiple transcription factors. Stable LSD1-knockdown cell lines were established by growing murine erythroleukemia (MEL) or mouse embryonic stem cells together with virus particles, in the presence of Polybrene® at 4 μg/mL, for 24–48 hours followed by puromycin selection (1 μg/mL) for 2 weeks. Real-time polymerase chain reaction (PCR)-based quantitative chromatin immunoprecipitation (ChIP) analysis was used to test whether the TAL1 transcription factor is bound to 1S promoter in the GATA-2 locus or whether LSD1 colocalizes with TAL1 at the 1S promoter. The sequential ChIP assay was utilized to confirm the role of LSD1 in the regulation of H3K4me2 at the GATA-2 locus during erythroid differentiation. Western blot analysis was employed to detect the protein expression. The alamarBlue® assay was used to examine the proliferation of the cells, and the absorbance was monitored at optical density (OD) 570 nm and OD 600 nm. In this study, we showed that LSD1 regulates the expression of GATA-2 during erythroid differentiation. Knockdown of LSD1 results in increased GATA-2 expression and inhibits the differentiation of MEL and embryonic stem cells. Furthermore, we demonstrated that LSD1 binds to the 1S promoter of the GATA-2 locus and suppresses GATA-2 expression, via histone demethylation. Our data revealed that LSD1 mediates erythroid differentiation, via epigenetic modification of the GATA-2 locus.