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Retinoids Induce Differential Expression and DNA Binding of the Mouse Germ Cell Nuclear Factor in P19 Embryonal Carcinoma Cells

机译:类维生素A诱导小鼠生殖细胞P19胚胎癌细胞中的差异表达和DNA结合。

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The mouse germ cell nuclear factor (GCNF), a member of the nuclear receptor superfamily, is highly expressed during gametogenesis and in the developing nervous system. The in vitro translated protein binds as a homodimer to the direct repeat (DR) of the sequence-AGGTCA- (DR-0). In this report, we characterize a DR-0 binding activity in P19 cell extracts that is induced by retinoids. This induction is concentration dependent and specific for embryonal carcinoma cells. The cellular protein binds with the same specificity as in vitro expressed GCNF, but migrates as a slower complex, indicating interaction with partner proteins. Because antisera directed against GCNF recognize this complex, we propose that GCNF is part of the binding activity. Combining in vitro translated GCNF and extracts of non-expressing cells shows that such interactions can be formed posttranslationally. Northern analysis demonstrates a concentration dependent induction of GCNF mRNA by retinoic acid. A time course shows that the level of GCNF binding is transiently elevated, later downregulated, and not detectable in differentiated cells. We propose that GCNF regulations is an important step during determination of embryonal carcinoma cells.
机译:小鼠生殖细胞核因子(GCNF)是核受体超家族的成员,在配子发生期间和发育中的神经系统中高度表达。体外翻译的蛋白质作为同源二聚体与序列AGGTCA-(DR-0)的直接重复序列(DR)结合。在此报告中,我们表征了类视色素诱导的P19细胞提取物中的DR-0结合活性。这种诱导是浓度依赖性的,并且对胚胎癌细胞具有特异性。细胞蛋白与体外表达的GCNF具有相同的特异性结合,但迁移速度较慢,表明与伴侣蛋白发生相互作用。因为针对GCNF的抗血清识别这种复合物,所以我们建议GCNF是结合活性的一部分。结合体外翻译的GCNF和非表达细胞的提取物表明,这种相互作用可以在翻译后形成。 Northern分析证明视黄酸对GCNF mRNA的浓度依赖性诱导。时间过程表明,GCNF结合水平瞬时升高,随后下调,并且在分化细胞中不可检测。我们建议,GCNF法规是确定胚胎癌细胞的重要步骤。

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