Drevet J R, Lareyre J J, Schwaab V, Vernet P, Dufaure J P
Laboratoire de Biologie Cellulaire, Université Blaise Pascal-Clermont Ferrand II, Aubière, France.
Mol Reprod Dev. 1998 Feb;49(2):131-40. doi: 10.1002/(SICI)1098-2795(199802)49:2<131::AID-MRD4>3.0.CO;2-Q.
This report presents data that suggest that the tissue-restricted polyoma enhancer activator protein (PEA3) of the Ets oncogene family of DNA-binding proteins is a putative modulator of the epididymis-specific glutathione peroxidase 5 gene gpx5. Northern and polymerase chain reactions on reverse-transcribed epididymal RNAs were used to show that the PEA3 factor is spatially and temporally expressed within the mouse epididymis in a manner consistent with gpx5 characteristics of expression. Then, using contransfection experiments carried out in heterologous tissue-culture cells with various deletions of the gpx5 promoter driving a CAT reporter gene, we have shown that the transcriptional activity of the gpx5 promoter is modulated by the presence of the PEA3 protein. Subsequently, we have shown using gel-shift assays that DNA sequences located within the 5' flanking region of the gpx5 gene have the ability to bind specifically to the PEA3 protein. Finally, using Northern assays we present data that suggest that PEA3 mRNA accumulation in the mouse caput epididymidis is controlled by androgens and testicular factors. Altogether, these results strongly suggest that the PEA3 factor might participate in the transcriptional control of the murine epididymis caput-specific gpx5 gene.
本报告呈现的数据表明，DNA结合蛋白Ets癌基因家族的组织限制性多瘤增强子激活蛋白（PEA3）可能是附睾特异性谷胱甘肽过氧化物酶5基因gpx5的调节因子。对逆转录的附睾RNA进行Northern印迹和聚合酶链反应，结果显示PEA3因子在小鼠附睾中呈现出与gpx5表达特征相一致的时空表达模式。然后，在异源组织培养细胞中进行共转染实验，使用携带不同缺失的gpx5启动子驱动CAT报告基因，结果表明gpx5启动子的转录活性受PEA3蛋白的存在调控。随后，通过凝胶迁移试验表明，gpx5基因5'侧翼区域内的DNA序列能够特异性结合PEA3蛋白。最后，利用Northern印迹分析，我们呈现的数据表明，小鼠附睾头中PEA3 mRNA的积累受雄激素和睾丸因子的控制。总之，这些结果有力地表明，PEA3因子可能参与了小鼠附睾头特异性gpx5基因的转录调控。
