Orford R L, Robinson C, Haydon J M, Patient R K, Guille M J
Biophysics Laboratories, Division of Molecular and Cell Biology, Institute of Biomolecular and Biomedical Sciences, University of Portsmouth, Portsmouth PO1 2DY, United Kingdom.
Mol Cell Biol. 1998 Sep;18(9):5557-66. doi: 10.1128/MCB.18.9.5557.
The transcription factor GATA-2 is expressed at high levels in the nonneural ectoderm of the Xenopus embryo at neurula stages, with lower amounts of RNA present in the ventral mesoderm and endoderm. The promoter of the GATA-2 gene contains an inverted CCAAT box conserved among Xenopus laevis, humans, chickens, and mice. We have shown that this sequence is essential for GATA-2 transcription during early development and that the factor binding it is maternal. The DNA-binding activity of this factor is detectable in nuclei and chromatin bound only when zygotic GATA-2 transcription starts. Here we report the characterization of this factor, which we call CBTF (CCAAT box transcription factor). CBTF activity mainly appears late in oogenesis, when it is nuclear, and the complex has multiple subunits. We have identified one subunit of the factor as p122, a Xenopus double-stranded-RNA-binding protein. The p122 protein is perinuclear during early embryonic development but moves from the cytoplasm into the nuclei of embryonic cells at stage 9, prior to the detection of CBTF activity in the nucleus. Thus, the accumulation of CBTF activity in the nucleus is a multistep process. We show that the p122 protein is expressed mainly in the ectoderm. Expression of p122 mRNA is more restricted, mainly to the anterior ectoderm and mesoderm and to the neural tube. Two properties of CBTF, its dual role and its cytoplasm-to-nucleus translocation, are shared with other vertebrate maternal transcription factors and may be general properties of these proteins.
转录因子GATA-2在非洲爪蟾胚胎神经胚阶段的非神经外胚层中高水平表达,在腹侧中胚层和内胚层中RNA含量较低。GATA-2基因的启动子包含一个反向CCAAT框,在非洲爪蟾、人类、鸡和小鼠中保守。我们已经表明,该序列对于早期发育过程中的GATA-2转录至关重要,并且与之结合的因子是母源性的。只有当合子GATA-2转录开始时,这种因子的DNA结合活性才能在仅与细胞核和染色质结合时检测到。在这里,我们报告了这种因子的特性,我们将其称为CBTF(CCAAT框转录因子)。CBTF活性主要在卵子发生后期出现,此时它位于细胞核中,并且该复合物有多个亚基。我们已经确定该因子的一个亚基为p122,一种非洲爪蟾双链RNA结合蛋白。p122蛋白在胚胎早期发育过程中位于核周,但在第9阶段从细胞质移入胚胎细胞的细胞核,此时细胞核中尚未检测到CBTF活性。因此,CBTF活性在细胞核中的积累是一个多步骤过程。我们表明p122蛋白主要在外胚层中表达。p122 mRNA的表达更具局限性,主要在前外胚层和中胚层以及神经管中。CBTF的两个特性,即其双重作用及其从细胞质到细胞核的易位,与其他脊椎动物母源性转录因子相同,可能是这些蛋白质的普遍特性。