Chromatin immunoprecipitation-mediated target identification proved aquaporin 5 is regulated directly by estrogen in the uterus
Chromatin immunoprecipitation-mediated target identification proved aquaporin 5 is regulated directly by estrogen in the uterus
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DOI：
10.1111/j.1365-2443.2006.01009.x
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发表时间：
2006-10-01
期刊：
影响因子：
2.1
通讯作者：
Iguchi, Taisen
中科院分区：
文献类型：
--
作者：
Kobayashi, Mika;Takahashi, Eri;Iguchi, Taisen
雌激素在脊椎动物的繁殖中发挥着核心作用，并影响多种生物过程。雌激素的主要靶分子是核雌激素受体（ER），已在分子水平上进行了广泛的研究。相比之下，我们对 ER 直接调节的基因的了解仍然有限，特别是在整个生物体而不是培养细胞的水平上。为了鉴定体内 ER 直接调节的基因，我们使用雌激素处理的小鼠子宫并进行染色质免疫沉淀。对沉淀的 DNA 片段进行序列分析，能够与小鼠基因组序列进行比对，并揭示编码水通道蛋白 5 (AQP5) 的基因的启动子区域是用抗 ER α 的抗体沉淀的。定量 PCR 和 DNA 微阵列分析证实，AQP5 在施用雌激素后很快被激活。此外，AQP5的启动子区含有功能性雌激素反应元件，可直接被雌激素激活。尽管一些 AQP 基因在子宫中表达，但在雌激素治疗后只能检测到 AQP5 的直接激活。这种染色质免疫沉淀介导的靶标识别可能适用于其他转录因子网络的研究。
Estrogens play a central role in the reproduction of vertebrates and affect a variety of biological processes. The major target molecules of estrogens are nuclear estrogen receptors (ERs), which have been studied extensively at the molecular level. In contrast, our knowledge of the genes that are regulated directly by ERs remains limited, especially at the level of the whole organism rather than cultured cells. In order to identify genes that are regulated directly by ERs in vivo, we used estrogen treated mouse uterus and performed chromatin immunoprecipitation. Sequence analysis of a precipitated DNA fragment enabled alignment with the mouse genomic sequence and revealed that the promoter region of the gene encoding aquaporin 5 (AQP5) was precipitated with antibody against ER alpha. Quantitative PCR and DNA microarray analyses confirmed that AQP5 is activated soon after administration of estrogen. In addition, the promoter region of AQP5 contained a functional estrogen response element that was activated directly by estrogen. Although several AQP genes are expressed in the uterus, only direct activation of AQP5 could be detected following treatment with estrogen. This chromatin immunopreciptation-mediated target identification may be applicable to the study of other transcription factor networks.