Cofactor CLIM2 promotes the repressive action of LIM homeodomain transcription factor Lhx2 in the expression of porcine pituitary glycoprotein hormone α subunit gene

Takao Susa, Takanobu Sato, Tetsuo Ono, Takako Kato, Yukio Kato
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引用次数: 13

Abstract

We have cloned a porcine orthologue of cofactor CLIM2 (Ldb1/NLI) from the porcine pituitary cDNA library by protein–protein interaction with the Yeast Two-Hybrid System using porcine Lhx2 as a bait protein. Porcine CLIM2 shows a high identity (99%) in the dimerization domain, nuclear localization signal and LIM binding domain with those of man and mouse. The expression of CLIM2 gene in the anterior pituitary lobe was detected during the porcine fetal and postnatal period by RT-PCR analysis, suggesting that this protein is constitutively expressing and plays a basic role in the anterior pituitary. Transfection assay to the pituitary tumor derived LβT2 cells, and the Chinese hamster ovary cells demonstrated that CLIM2 acts as a corepressor of the porcine Lhx2 function. Interestingly, CLIM2 alone apparently repressed the high level of αGSU gene expression in LβT2 cells. These data suggest that CLIM2 is a basic factor in the pituitary development and function, and plays the role of repressor to modify the function of Lhx2 on the αGSU gene expression.

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辅助因子CLIM2促进LIM同源结构域转录因子Lhx2对猪垂体糖蛋白激素α亚基基因表达的抑制作用
以猪Lhx2为诱饵蛋白,利用酵母双杂交系统蛋白-蛋白相互作用,从猪垂体cDNA文库中克隆出猪辅助因子CLIM2的同源物(Ldb1/NLI)。猪的CLIM2在二聚化结构域、核定位信号和LIM结合结构域上与人和小鼠具有高度的一致性(99%)。RT-PCR检测了猪胎儿和产后垂体前叶中CLIM2基因的表达,表明该蛋白在垂体前叶中具有组成性表达,在垂体前叶中起基础作用。对垂体瘤源性LβT2细胞和中国仓鼠卵巢细胞的转染实验表明,CLIM2可作为猪Lhx2功能的协同抑制因子。有趣的是,单独使用CLIM2可以明显抑制LβT2细胞中α - gsu基因的高水平表达。这些数据表明,CLIM2是垂体发育和功能的基础因子,并发挥抑制因子的作用,调节Lhx2对αGSU基因表达的功能。
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