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首页> 外文期刊>Molecular and Cellular Biology >Characterization of a Multisubunit Transcription Factor Complex Essential for Spliced-Leader RNA Gene Transcription in Trypanosoma brucei
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Characterization of a Multisubunit Transcription Factor Complex Essential for Spliced-Leader RNA Gene Transcription in Trypanosoma brucei

机译：布鲁氏锥虫的拼接领导RNA基因转录所必需的多亚基转录因子复合物的表征。

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In the unicellular human parasites Trypanosoma brucei, Trypanosoma cruzi, and Leishmania spp., the spliced-leader (SL) RNA is a key molecule in gene expression donating its 5′-terminal region in SL addition trans splicing of nuclear pre-mRNA. While there is no evidence that this process exists in mammals, it is obligatory in mRNA maturation of trypanosomatid parasites. Hence, throughout their life cycle, these organisms crucially depend on high levels of SL RNA synthesis. As putative SL RNA gene transcription factors, a partially characterized small nuclear RNA-activating protein complex (SNAPc) and the TATA-binding protein related factor 4 (TRF4) have been identified thus far. Here, by tagging TRF4 with a novel epitope combination termed PTP, we tandem affinity purified from crude T. brucei extracts a stable and transcriptionally active complex of six proteins. Besides TRF4 these were identified as extremely divergent subunits of SNAPc and of transcription factor IIA (TFIIA). The latter finding was unexpected since genome databases of trypanosomatid parasites appeared to lack general class II transcription factors. As we demonstrate, the TRF4/SNAPc/TFIIA complex binds specifically to the SL RNA gene promoter upstream sequence element and is absolutely essential for SL RNA gene transcription in vitro.
机译：在单细胞人类寄生虫 Trypanosoma brucei ， Trypanosoma cruzi 和 Leishmania spp。中，剪接前导（SL）RNA是一个重要的分子。基因表达在核前体mRNA的SL加成反转录剪接中捐赠了其5'-末端区域。尽管没有证据表明该过程存在于哺乳动物中，但它在锥虫寄生虫的mRNA成熟中是必须的。因此，在整个生命周期中，这些生物至关重要地依赖于高水平的SL RNA合成。作为推定的SL RNA基因转录因子，到目前为止，已鉴定出部分表征的小核RNA活化蛋白复合物（SNAP c ）和TATA结合蛋白相关因子4（TRF4）。在这里，通过用称为PTP的新型表位组合标记TRF4，我们可以串联从粗Tem中纯化的亲和力。 brucei 提取出一种稳定且具有转录活性的六种蛋白质复合物。除TRF4外，它们还被确定为SNAP c 和转录因子IIA（TFIIA）的极其不同的亚基。后一种发现是出乎意料的，因为锥虫的寄生虫的基因组数据库似乎缺乏一般的II类转录因子。正如我们所证明的，TRF4 / SNAP c / TFIIA复合物特异性结合SL RNA基因启动子上游序列元件，并且对于SL RNA基因体外转录绝对必要。

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