Han SangYoon, Xie Wensheng, Hammes Stephen R, DeJong Jeff
Department of Molecular and Cell Biology, University of Texas at Dallas, Richardson, Texas 75080, USA.
J Biol Chem. 2003 Nov 14;278(46):45586-93. doi: 10.1074/jbc.M302884200. Epub 2003 Aug 14.
The discovery of germ cell-specific general transcription factor and coactivator variants has suggested that reproductive tissues control gene expression somewhat differently than somatic tissues. One of these factors, ALF (TFIIAtau), was first described as a testis-specific counterpart of the large (alpha/beta) subunit of TFIIA. Here we characterize endogenous ALF and TFIIA activities in the African clawed frog Xenopus laevis. ALF is present in both testis and ovary in this organism, and it completely replaces TFIIA in immature oocytes. When oocytes undergo progesterone-induced maturation, ALF activity disappears, and TFIIA activity is restored. Reactivation occurs through the translational up-regulation of two maternal TFIIAalpha/beta mRNAs and involves polyadenylation of a conserved 3'-untranslated region module. The effects of ALF overexpression and ALF immunodepletion on a thymidine kinase promoter construct demonstrate that this factor serves as an active replacement for TFIIA. In contrast, overexpression of TFIIA inhibits transcription, indicating that the somatic factor fails to function properly in the context of the oocyte transcription machinery. Overall, the results show that the translationally regulated reciprocal expression of ALF and TFIIA allows for the production of an active TFIIA-like general transcription factor throughout oogenesis.
生殖细胞特异性通用转录因子和共激活因子变体的发现表明,生殖组织控制基因表达的方式与体细胞组织有所不同。其中一个因子ALF(TFIIAtau)最初被描述为TFIIA大亚基(α/β)的睾丸特异性对应物。在这里,我们对非洲爪蟾(非洲爪蟾)体内的内源性ALF和TFIIA活性进行了表征。在这种生物体中,ALF存在于睾丸和卵巢中,并且它在未成熟卵母细胞中完全取代了TFIIA。当卵母细胞经历孕酮诱导的成熟过程时,ALF活性消失,TFIIA活性恢复。重新激活是通过两种母源TFIIAα/β mRNA的翻译上调发生的,并且涉及一个保守的3'非翻译区模块的多聚腺苷酸化。ALF过表达和ALF免疫耗竭对胸苷激酶启动子构建体的影响表明,该因子可作为TFIIA的活性替代物。相比之下,TFIIA的过表达会抑制转录,这表明体细胞因子在卵母细胞转录机制的背景下无法正常发挥作用。总体而言,结果表明,ALF和TFIIA的翻译调控的相互表达使得在整个卵子发生过程中能够产生一种活性的TFIIA样通用转录因子。