Joos T O, David R, Dreyer C
Max-Planck-Institut für Entwicklungsbiologie, Tübingen, Germany.
Mech Dev. 1996 Nov;60(1):45-57. doi: 10.1016/s0925-4773(96)00599-0.
Nuclear orphan receptors are DNA binding proteins that share the domain structure of the nuclear hormone receptor superfamily, although ligands are unknown. We have identified an orphan receptor in Xenopus laevis and named it xGCNF based on its high degree of sequence homology to the previously described murine germ cell nuclear factor (mGCNF). In gel-electrophoresis mobility shift analysis experiments in vitro translated xGCNF and mGCNF proteins both bind specifically as homodimers to the same response element, a direct repeat of the half-site consensus AGGTCA with zero spacing (DRO). Transcripts of xGCNF are found in oocytes and in much smaller amounts in the testes. In developmental Northern blots and RNase protection using RNA from different embryonic stages, zygotic expression of xGCNF peaks at midneurula. From late gastrula to midneurula stages, an anterior to posterior concentration gradient of the RNA was observed in whole mount in situ analysis. This antero-posterior gradient of expression was also observed in exogastrulae, both in the ectoderm and mesoderm. In the midneurula embryo, the mRNA was predominantly found in the neural plate and neural crest. Transcription of xGCNF in animal cap explants occurred independent of mesoderm induction.
核孤儿受体是一类DNA结合蛋白,它们具有核激素受体超家族的结构域结构,尽管其配体尚不清楚。我们在非洲爪蟾中鉴定出一种孤儿受体,并根据其与先前描述的小鼠生殖细胞核因子(mGCNF)的高度序列同源性将其命名为xGCNF。在凝胶电泳迁移率变动分析实验中,体外翻译的xGCNF和mGCNF蛋白均以同二聚体形式特异性结合至相同的反应元件,即半位点共有序列AGGTCA的零间隔直接重复序列(DRO)。xGCNF的转录本在卵母细胞中存在,在睾丸中的含量则少得多。在使用不同胚胎阶段的RNA进行的发育Northern印迹和RNA酶保护实验中,xGCNF的合子表达在神经中胚层期达到峰值。在原肠胚晚期至神经中胚层期阶段,在整体原位分析中观察到RNA从前向后的浓度梯度。在外胚层发育不全的胚胎中,在外胚层和中胚层中也观察到了这种前后表达梯度。在神经中胚层期胚胎中,mRNA主要存在于神经板和神经嵴中。xGCNF在动物帽外植体中的转录独立于中胚层诱导发生。