Pfaff S L, Hall R K, Hart G C, Taylor W L
School of Medicine, Department of Molecular Physiology and Biophysics, Vanderbilt University, Nashville, Tennessee 37232.
Dev Biol. 1991 Jun;145(2):241-54. doi: 10.1016/0012-1606(91)90123-k.
Expression of the Xenopus laevis transcription factor IIIA (TFIIIA) gene is developmentally regulated. In this study we have used defined nucleotide mutations to map cis-elements involved in transcriptional regulation of the promoter for oocyte-TFIIIA (O-TFIIIA) in stage II-IV oocytes, stage VI oocytes, and tail bud embryos. During oogenesis O-TFIIIA mRNA levels decline 5- to 10-fold, and during early embryogenesis O-TFIIIA mRNA levels decline approximately 10(6)-fold per cell. In stage II-IV oocytes we find evidence for at least three distinct positive-acting cis-elements that contribute to the efficient expression of O-TFIIIA. These elements are located between -1800 to -425, -280 to -235, and -235 to -220. The most distal cis-element(s) appears to be developmentally regulated during oogenesis, since deletion of nucleotide sequences from -1800 to -425 does not reduce O-TFIIIA expression in stage VI oocytes. However, the two cis-elements located between -280 to -235 and -235 to -220 are required for the efficient expression of O-TFIIIA in stage VI oocytes. In tail bud embryos we find evidence for several developmentally regulated positive and negative cis-elements involved in O-TFIIIA expression. The positive-acting cis-elements are located between -159 to -110 and -110 to -58. The negative-acting cis-elements are found at positions -425 to -350 and -200 to -159. In addition to the developmentally regulated elements controlling O-TFIIIA gene expression in tail bud embryos, the positive-acting cis-elements active during oogenesis (located between -280 to -235 and -235 to -220) are also active during early embryogenesis. Thus, transcription from the O-TFIIIA promoter appears to be regulated by a combination of constitutive positive factors and developmentally regulated positive and negative factors during oogenesis and early embryogenesis.
非洲爪蟾转录因子IIIA(TFIIIA)基因的表达受到发育调控。在本研究中,我们利用特定的核苷酸突变来定位参与卵母细胞TFIIIA(O-TFIIIA)启动子转录调控的顺式元件,该启动子存在于II-IV期卵母细胞、VI期卵母细胞和尾芽胚胎中。在卵子发生过程中,O-TFIIIA mRNA水平下降5至10倍,而在早期胚胎发生过程中,每个细胞中O-TFIIIA mRNA水平下降约10⁶倍。在II-IV期卵母细胞中,我们发现至少有三个不同的正向作用顺式元件有助于O-TFIIIA的高效表达。这些元件位于-1800至-425、-280至-235和-235至-220之间。最远端的顺式元件在卵子发生过程中似乎受到发育调控,因为从-1800至-425删除核苷酸序列并不会降低VI期卵母细胞中O-TFIIIA的表达。然而,位于-280至-235和-235至-220之间的两个顺式元件是VI期卵母细胞中O-TFIIIA高效表达所必需的。在尾芽胚胎中,我们发现了几个参与O-TFIIIA表达的发育调控的正向和负向顺式元件。正向作用的顺式元件位于-159至-110和-110至-58之间。负向作用的顺式元件位于-425至-350和-200至-159位置。除了在尾芽胚胎中控制O-TFIIIA基因表达的发育调控元件外,卵子发生过程中活跃的正向作用顺式元件(位于-280至-235和-235至-220之间)在早期胚胎发生过程中也活跃。因此,在卵子发生和早期胚胎发生过程中,O-TFIIIA启动子的转录似乎受组成型正向因子以及发育调控型正向和负向因子的组合调控。