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首页> 外文期刊>Journal of Molecular Biology >REPRESSION OF VERTEBRATE RNA POLYMERASE III TRANSCRIPTION BY DNA BINDING PROTEINS LOCATED UPSTREAM FROM THE TRANSCRIPTION START SITE
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REPRESSION OF VERTEBRATE RNA POLYMERASE III TRANSCRIPTION BY DNA BINDING PROTEINS LOCATED UPSTREAM FROM THE TRANSCRIPTION START SITE

机译：从转录起始位点定位的DNA结合蛋白抑制VERTERBRATE RNA聚合酶III的转录

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Derivatives of yeast tRNA and Xenopus tRNA and 5 S RNA genes have been constructed in which natural 5' flanking sequences have been replaced by the binding sites for either the yeast transcription activator protein GCN4 or the three amino-terminal zinc fingers of the Xenopus factor TFIIIA (zf1-3). The binding sites for these proteins have been placed at various distances upstream from the start site for transcription initiation in the parent genes. Each of these plasmid DNAs is actively transcribed in both an unfractionated transcription extract prepared from unfertilized Xenopus eggs and in a reconstituted Xenopus transcription system. Binding of the test proteins to plasmid DNAs harboring the cognate binding sites severely represses transcription when these binding sites are located less than approximately 40 base-pairs upstream from the transcription start site. The DNA-binding proteins are without effect on the transcription of plasmids lacking binding sites or when the binding sites are located further upstream. Assembly of DNA templates into a complete transcription complex prior to addition of the DNA-binding proteins prevents repression. Proteins present in a fraction containing TFIIIB are necessary for this reversal of repression. These data suggest that vertebrate TFIIIB binds upstream from class III genes and this binding can be prevented by occlusion of the TFIIIB binding site by the test proteins GCN4 and zf1-3. [References: 40]
机译：已构建酵母tRNA和非洲爪蟾tRNA和5 S RNA基因的衍生物，其中天然5'侧翼序列已被酵母转录激活蛋白GCN4或非洲爪蟾因子TFIIIA的三个氨基末端锌指的结合位点取代（zf1-3）。这些蛋白的结合位点已被放置在起始位点上游不同距离处，以在亲本基因中进行转录起始。这些质粒DNA中的每一个都在从未受精的爪蟾卵制备的未分级转录提取物中和在重组的爪蟾转录系统中被主动转录。当这些结合位点位于转录起始位点上游少于约40个碱基对时，测试蛋白质与具有同源结合位点的质粒DNA的结合会严重抑制转录。 DNA结合蛋白对缺乏结合位点的质粒的转录没有影响，或者当结合位点位于更上游时没有影响。在添加DNA结合蛋白之前，将DNA模板组装成完整的转录复合物可防止阻抑。存在于包含TFIIIB的级分中的蛋白质对于这种逆转抑制是必需的。这些数据表明，脊椎动物TFIIIB在III类基因的上游结合，并且可以通过测试蛋白GCN4和zf1-3封闭TFIIIB结合位点来防止这种结合。 [参考：40]

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