Estrogen receptor β exon 3-deleted mouse: The importance of non-ERE pathways in ERβ signaling
Estrogen receptor β exon 3-deleted mouse: The importance of non-ERE pathways in ERβ signaling
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DOI：
10.1073/pnas.1504944112
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发表时间：
2015-04-21
影响因子：
11.1
通讯作者：
Gustafsson, Jan-Ake
中科院分区：
文献类型：
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作者：
Maneix, Laure;Antonson, Per;Gustafsson, Jan-Ake
在1998年，通过用新盒在DNA结合域（DBD）处中断基因来创建雌激素受体β（ER β）敲除（KO）小鼠。突变的雌性动物生育能力低下，大脑、前列腺、肺、结肠和免疫系统都有异常。在2008年，另一种ER β突变小鼠是通过缺失编码DBD中第一个锌指的ER β外显子3产生的。该品系的雌性小鼠不能排卵，但其他方面正常。两种KO菌株表型的差异导致了关于ER β生理功能的问题。在本研究中，我们创建了一个ER β外显子3缺失的小鼠（ER β-Delta ex 3），并证实唯一可观察到的缺陷是无排卵。尽管通过外显子2和4之间的剪接引入了两个框内终止密码子，但ER β蛋白在前列腺上皮细胞的细胞核中表达。使用两种不同的抗ER β抗体，我们发现ER β-Delta ex 3蛋白中存在框内配体结合结构域和C末端。此外，从ER β-Delta ex 3前列腺细胞核提取物，有一个ER β依赖性阻滞迁移的激活蛋白-1的反应元件在EMSA。与原始基因敲除小鼠不同，ER β-Delta ex 3小鼠前列腺上皮中Ki 67、雄激素受体和Dachshund-1的表达没有改变。我们的结论是，很少的ER β转录活性依赖于结合经典的雌激素反应元件（ERE）。
In 1998, an estrogen receptor beta (ER beta) knockout (KO) mouse was created by interrupting the gene at the DNA binding domain (DBD) with a neocassette. The mutant females were subfertile and there were abnormalities in the brain, prostate, lung, colon, and immune system. In 2008, another ER beta mutant mouse was generated by deleting ER beta exon 3 which encodes the first zinc finger in the DBD. The female mice of this strain were unable to ovulate but were otherwise normal. The differences in the phenotypes of the two KO strains, have led to questions about the physiological function of ER beta. In the present study, we created an ER beta exon 3-deleted mouse (ER beta-Delta ex3) and confirmed that the only observable defect was anovulation. Despite the two in-frame stop codons introduced by splicing between exons 2 and 4, an ER beta protein was expressed in nuclei of prostate epithelial cells. Using two different anti-ER beta antibodies, we showed that an in-frame ligand binding domain and C terminus were present in the ER beta-Delta ex3 protein. Moreover, with nuclear extracts from ER beta-Delta ex3 prostates, there was an ER beta-dependent retardation of migration of activator protein-1 response elements in EMSA. Unlike the original knockout mouse, expression of Ki67, androgen receptor, and Dachshund-1 in prostate epithelium was not altered in the ER beta-Delta ex3 mouse. We conclude that very little of ER beta transcriptional activity depends on binding to classical estrogen response elements (EREs).