Distinct roles for CBP and p300 on the RA-mediated expression of the meiosis commitment gene Stra8 in mouse embryonic stem cells.
Distinct roles for CBP and p300 on the RA-mediated expression of the meiosis commitment gene Stra8 in mouse embryonic stem cells.
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CBP 和 p300 对 RA 介导的小鼠胚胎干细胞减数分裂基因 Stra8 表达的不同作用
DOI：
10.1371/journal.pone.0066076
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Kang J
中科院分区：
文献类型：
--
作者：
Chen W;Jia W;Wang K;Si X;Zhu S;Duan T;Kang J
在哺乳动物生殖细胞中，减数分裂定型需要视黄酸刺激基因8（Stra8）的表达，其被视黄酸（RA）转录激活。然而，鲜为人知的是，RA诱导Stra8表达的表观遗传机制。利用染色质免疫沉淀试验（ChIP），我们发现，RA增加组蛋白乙酰化在Stra8启动子在小鼠胚胎干细胞（ESCs），生殖细胞分化的模型。此外，我们探讨了两个与组蛋白乙酰转移酶（HAT）活性，Creb结合蛋白（CBP）和p300，是否参与激活Stra8。内源性CBP的慢病毒shRNA敲低导致Stra8抑制，而CBP的过表达在mRNA和蛋白水平上增强Stra8表达。ChIP分析证实CBP是RA介导的Stra8转录的关键共激活因子，它增强了组蛋白乙酰化水平，并招募RNA聚合酶II以建立转录活性染色质。此外，p300的shRNA增强了Stra8的表达，而p300的过表达降低了Stra8的表达，这与其HAT活性无关。ChIP显示敲除p300显著增加了Stra8启动子处的CBP水平。这些发现表明CBP和p300在RA介导的Stra8基因转录中发挥不同的作用。
In mammalian germ cells, meiotic commitment requires the expression of Stimulated by retinoic acid gene 8 (Stra8), which is transcriptionally activated by retinoic acid (RA). However, little is known about the epigenetic mechanism by which RA induces Stra8 expression. Utilizing a chromatin immunoprecipitation assay (ChIP), we showed that RA increases histone acetylation at the Stra8 promoter in murine embryonic stem cells (ESCs), a model for germ cell differentiation. Furthermore, we explored whether two coregulators with histone acetyltransferase (HAT) activity, Creb-binding protein (CBP) and p300, are involved in the activation of Stra8. The lentiviral shRNA knockdown of endogenous CBP led to Stra8 repression, while the overexpression of CBP enhanced Stra8 expression at both the mRNA and protein levels. ChIP analysis confirmed that CBP is the crucial coactivator for RA-mediated Stra8 transcription and that it enhances the level of histone acetylation and recruits RNA polymerase II to establish transcriptionally active chromatin. Furthermore, shRNA of p300 enhanced Stra8 expression, and the overexpression of p300 reduced Stra8 expression, independently of its HAT activity. ChIP showed that the knockdown of p300 significantly increased the level of CBP at the Stra8 promoter. These findings demonstrate that CBP and p300 play distinct roles in RA-mediated Stra8 gene transcription.
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