Ada3 requirement for HAT recruitment to estrogen receptors and estrogen-dependent breast cancer cell proliferation
Ada3 requirement for HAT recruitment to estrogen receptors and estrogen-dependent breast cancer cell proliferation
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DOI：
10.1158/0008-5472.can-07-2721
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发表时间：
2007-12-15
期刊：
影响因子：
11.2
通讯作者：
Band, Vimla
中科院分区：
文献类型：
--
作者：
Germaniuk-Kurowska, Aleksandra;Nag, Alo;Band, Vimla
我们以前已经表明，进化保守的激活（Ada）蛋白的改变/缺陷与雌激素受体（ER）介导的靶基因的表达，并促进。在这里，我们研究了内源性Ada 3的作用，招募组蛋白乙酰转移酶（RAT）的ER-响应启动子和雌激素依赖性细胞增殖和恶性表型的作用。使用甘油梯度共沉淀和免疫沉淀分析的组合，我们表明，Ada 3，ER，和三个不同的HAT [p300，（p300/CBP相关因子）PCAF，和一般控制nonrepressed 5（Gcn 5）]存在于一个复杂的。使用染色质免疫沉淀分析，我们表明，短发夹RNA（shRNA）介导的敲低雌激素受体阳性乳腺癌细胞中的Ada 3显着减少了配体依赖性招聘的p300，PCAF，和Gcn 5的ER-响应的pS2启动子。最后，我们使用shRNA敲低表明，Ada 3是关键的雌激素依赖性增殖的ER阳性乳腺癌细胞系在二维，以及三维，文化。在ER阳性MCF-7细胞中敲低Ada 3诱导三维培养中转化表型的逆转。因此，我们的研究结果表明，Ada 3在HAT招募雌激素反应性靶基因启动子和乳腺癌细胞的雌激素依赖性增殖中起重要作用。
We have previously shown that evolutionarily conserved alteration/deficiency in activation (Ada) protein associates with and promotes estrogen receptor (ER)-mediated target gene expression. Here, we examined the role of endogenous Ada3 to recruit histone acetyl transferases (RAT) to an ER-responsive promoter and its role in estrogen-dependent cell proliferation and malignant phenotype. Using a combination of glycerol gradient cosedimentation and immunoprecipitation analyses, we show that Ada3, ER, and three distinct HATs [p300, (p300/CBP-associated factor) PCAF, and general control nonrepressed 5 (Gcn5)] are present in a complex. Using chromatin immunoprecipitation analysis, we show that short hairpin RNA (shRNA) -mediated knockdown of Ada3 in ER-positive breast cancer cells significantly reduced the ligand-dependent recruitment of p300, PCAF, and Gcn5 to the ER-responsive pS2 promoter. Finally, we use shRNA knockdown to show that Ada3 is critical for estrogen-dependent proliferation of ER-positive breast cancer cell lines in two-dimensional, as well as three-dimensional, culture. Knockdown of Ada3 in ER-positive MCF-7 cells induced reversion of the transformed phenotype in three-dimensional culture. Thus, our results show an important role of Ada3 in HAT recruitment to estrogen-responsive target gene promoters and for estrogen-dependent proliferation of breast cancer cells.