Lawinger P, Rastelli L, Zhao Z, Majumder S
University of Texas M. D. Anderson Cancer Center, Houston, Texas 77030, USA.
J Biol Chem. 1999 Mar 19;274(12):8002-11. doi: 10.1074/jbc.274.12.8002.
Previous studies have shown that the lack of novel coactivator activity in mouse oocytes and one-cell embryos (fertilized eggs) renders them incapable of utilizing Gal4:VP16-dependent enhancers (distal elements) but not promoters (proximal elements) in regulating transcription. This coactivator activity first appears in two- to four-cell embryos coincident with the major activation of zygotic gene expression. Here we show that whereas oocytes and fertilized eggs could utilize Sp1-dependent promoters, they could not utilize Sp1-dependent enhancers, although they showed promoter repression, which is a requirement for delineating enhancer function. In contrast, both Sp1-dependent promoters and enhancers were functional in two- to four-cell embryos. Furthermore, the same embryonic stem cell mRNA that provided the coactivator activity for Gal4:VP16-dependent enhancer function also provided Sp1-dependent enhancer function in oocytes. Therefore, the coactivator activity appears to be a requirement for general enhancer function. To determine whether the absence of enhancer function is a unique property of oocytes or a general property of other terminally differentiated cells, transcription was examined in terminally differentiated hNT neurons and their precursors, undifferentiated NT2 stem cells. The results showed that both cell types could utilize enhancers and promoters. Thus, in mammals, the lack of enhancer function appears to be unique to oocytes and fertilized eggs, suggesting that it provides a safeguard against premature activation of genes prior to zygotic gene expression during development.
先前的研究表明,小鼠卵母细胞和单细胞胚胎(受精卵)缺乏新型共激活因子活性,这使得它们无法利用Gal4:VP16依赖性增强子(远端元件)来调控转录,但在调控转录时能够利用启动子(近端元件)。这种共激活因子活性首先出现在二细胞至四细胞胚胎中,与合子基因表达的主要激活同时发生。在这里,我们表明,虽然卵母细胞和受精卵可以利用Sp1依赖性启动子,但它们不能利用Sp1依赖性增强子,尽管它们表现出启动子抑制,这是界定增强子功能的一个必要条件。相比之下,Sp1依赖性启动子和增强子在二细胞至四细胞胚胎中均具有功能。此外,为Gal4:VP16依赖性增强子功能提供共激活因子活性的同一胚胎干细胞mRNA,在卵母细胞中也为Sp1依赖性增强子功能提供了支持。因此,共激活因子活性似乎是一般增强子功能所必需的。为了确定增强子功能的缺失是卵母细胞的独特特性还是其他终末分化细胞的普遍特性,我们对终末分化的hNT神经元及其前体、未分化的NT2干细胞进行了转录检测。结果表明,这两种细胞类型都可以利用增强子和启动子。因此,在哺乳动物中,增强子功能的缺失似乎是卵母细胞和受精卵所特有的,这表明它为发育过程中合子基因表达之前的基因过早激活提供了一种保护机制。