Tsai-Pflugfelder M, Gasser S M, Wahli W
Institut de Biologie animale, Université de Lausanne, Switzerland.
Mol Endocrinol. 1998 Oct;12(10):1525-41. doi: 10.1210/mend.12.10.0182.
Eukaryotic gene expression depends on a complex interplay between the transcriptional apparatus and chromatin structure. We report here a yeast model system for investigating the functional interaction between the human estrogen receptor (hER) and CTF1, a member of the CTF/NFI transcription factor family. We show that a CTF1-fusion protein and the hER transactivate a synthetic promoter in yeast in a synergistic manner. This interaction requires the proline-rich transactivation domain of CTF1. When the natural estrogen-dependent vitellogenin B1 promoter is tested in yeast, CTF1 and CTF1-fusion proteins are unable to activate transcription, and no synergy is observed between hER, which activates the B1 promoter, and these factors. Chromatin structure analysis on this promoter reveals positioned nucleosomes at -430 to -270 (+/-20 bp) and at -270 to - 100 (+/-20 bp) relative to the start site of transcription. The positions of the nucleosomes remain unchanged upon hormone-dependent transcriptional activation of the promoter, and the more proximal nucleosome appears to mask the CTF/NFI site located at - 101 to -114. We conclude that a functional interaction of hER with the estrogen response element located upstream of a basal promoter occurs in yeast despite the nucleosomal organization of this promoter, whereas the interaction of CTF1 with its target site is apparently precluded by a nucleosome.
真核基因表达取决于转录装置与染色质结构之间复杂的相互作用。我们在此报告一个酵母模型系统,用于研究人类雌激素受体(hER)与CTF1(CTF/NFI转录因子家族成员之一)之间的功能相互作用。我们发现CTF1融合蛋白与hER以协同方式在酵母中激活一个合成启动子。这种相互作用需要CTF1富含脯氨酸的转录激活结构域。当在酵母中测试天然雌激素依赖性卵黄蛋白原B1启动子时,CTF1和CTF1融合蛋白无法激活转录,并且在激活B1启动子的hER与这些因子之间未观察到协同作用。对该启动子的染色质结构分析显示,相对于转录起始位点,在-430至-270(±20 bp)以及-270至-100(±20 bp)处存在定位核小体。在启动子的激素依赖性转录激活过程中,核小体的位置保持不变,并且更靠近近端的核小体似乎掩盖了位于-101至-114的CTF/NFI位点。我们得出结论,尽管该启动子存在核小体组织,但hER与位于基础启动子上游的雌激素反应元件在酵母中仍发生功能相互作用,而CTF1与其靶位点的相互作用显然被一个核小体所阻止。