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首页> 外文期刊>Molecular and Cellular Biology >Robust mRNA Transcription in Chicken DT40 Cells Depleted of TAFII31 Suggests Both Functional Degeneracy and Evolutionary Divergence
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Robust mRNA Transcription in Chicken DT40 Cells Depleted of TAFII31 Suggests Both Functional Degeneracy and Evolutionary Divergence

机译：TAFII31耗尽的鸡DT40细胞中的稳健mRNA转录表明功能简并性和进化差异

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摘要

We have employed gene targeting coupled with conditional expression to construct a chicken DT40 cell line in which a tetracycline (Tet)-repressible promoter is exclusively responsible for expression of cTAFII31, a histone-like TAFII residing in both the transcription factor TFIID and the histone acetylase complex PCAF/SAGA. Tet addition resulted in rapid loss of cTAFII31 mRNA and protein, eventually leading to apoptotic cell death. Significantly, five of six other TAFIIs tested were also rapidly depleted, but levels of the TATA binding protein and subunits of PCAF/SAGA were at most modestly compromised. Strikingly, pulse-labeling experiments indicate that total poly(A)+mRNA transcription was not significantly reduced after cTAFII31 depletion, and steady-state levels of several specific transcripts remained the same or decreased only mildly. Moreover, activation of c-fos transcription following serum starvation occurred efficiently in the absence of cTAFII31. These data, which contrast with comparable studies in yeast, strongly suggest that cTAFII31 and perhaps other TAFIIs are not essential for general mRNA transcription in DT40 cells. We propose that this is due to extensive functional degeneracy in the highly complex metazoan transcriptional machinery.
机译：我们已经利用基因靶向结合条件表达来构建鸡DT40细胞系，其中四环素（Tet）可阻遏的启动子专门负责表达cTAF II 31（组蛋白样TAF > II 同时存在于转录因子TFIID和组蛋白乙酰化酶复合物PCAF / SAGA中。 Tet的加入导致cTAF II 31 mRNA和蛋白迅速丢失，最终导致凋亡细胞死亡。值得注意的是，其他六种TAF II 中的五种也迅速耗尽，但是TATA结合蛋白和PCAF / SAGA亚基的水平受到的损害最大。令人惊讶的是，脉冲标记实验表明，cTAF II 31耗尽后，poly（A） + mRNA的总转录没有显着降低，并且仍保留了几种特定转录本的稳态水平相同或仅轻微下降。此外，在缺乏cTAF II 31的情况下，血清饥饿后c- fos 转录的激活有效发生。这些数据与酵母中的可比研究形成对比，强烈表明cTAF II 31和其他TAF II 并不是DT40细胞中一般mRNA转录所必需的。我们建议这是由于高度复杂的后生动物转录机制中广泛的功能简并性。

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