Brewer A C, Guille M J, Fear D J, Partington G A, Patient R K
Developmental Biology Research Centre, Randall Institute, King's College, London, UK.
EMBO J. 1995 Feb 15;14(4):757-66. doi: 10.1002/j.1460-2075.1995.tb07054.x.
The transcription factor GATA-2 is present in blood cell precursors and plays a pivotal role in the control of erythroid differentiation. In Xenopus embryos, low levels of GATA-2 mRNA are maternally derived, while the onset of zygotic GATA-2 expression coincides with commitment to haematopoietic lineages. However, its initial transcriptional activation is not restricted to the presumptive blood islands, but occurs throughout ventral and lateral regions, in all three germ layers. In order to determine how this expression pattern is controlled, we have isolated and characterized the Xenopus GATA-2 gene. We show that 1.65 kb of 5' flanking sequences are sufficient to direct both correct transcriptional initiation in oocytes and appropriate temporal and spatial gene expression in early embryos. The transgene is activated during gastrulation and by neurula stages in predominantly expressed in the ventral hemisphere. We demonstrate that a CCAAT element is necessary for gene activity in both systems and that extracts prepared from oocytes and embryos contain a factor which specifically recognizes this element. We also show that cytoplasmic localization inhibits the function of this CCAAT factor until the beginning of gastrulation, when the zygotic GATA-2 gene is activated. These observations extend our understanding of the mechanisms by which maternal factors control the temporal activation of transcription in early vertebrate embryos.
转录因子GATA - 2存在于血细胞前体中,在红细胞分化的调控中起关键作用。在非洲爪蟾胚胎中,低水平的GATA - 2 mRNA是母源的,而合子型GATA - 2表达的起始与造血谱系的确定相吻合。然而,其最初的转录激活并不局限于假定的血岛,而是发生在所有三个胚层的腹侧和外侧区域。为了确定这种表达模式是如何被调控的,我们分离并鉴定了非洲爪蟾GATA - 2基因。我们发现1.65 kb的5'侧翼序列足以指导卵母细胞中的正确转录起始以及早期胚胎中适当的时空基因表达。转基因在原肠胚形成期间被激活,到神经胚阶段主要在腹侧半球表达。我们证明CCAAT元件对于两个系统中的基因活性都是必需的,并且从卵母细胞和胚胎中制备的提取物含有一种能特异性识别该元件的因子。我们还表明,细胞质定位会抑制该CCAAT因子的功能,直到原肠胚形成开始时合子型GATA - 2基因被激活。这些观察结果扩展了我们对母源因子控制早期脊椎动物胚胎转录时间激活机制的理解。