Evolution of p53 transactivation specificity through the lens of a yeast-based functional assay.
Evolution of p53 transactivation specificity through the lens of a yeast-based functional assay.
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DOI：
10.1371/journal.pone.0116177
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Inga A
中科院分区：
文献类型：
--
作者：
Lion M;Raimondi I;Donati S;Jousson O;Ciribilli Y;Inga A
转录因子与其各自的顺式调控网络的共同进化增强了进化过程中的功能多样性。我们提出了一种新的方法来研究序列特异性转录因子在进化研究中的反式激活能力。以酿酒酵母为体内试管，选择来自人类和5种常用动物模型的P53蛋白作为概念验证。P53是一种高度保守的环境应激反应的主要调节因子。以往的报道表明，P53 DNA在体外具有保守的结合特异性，即使是在进化较远的物种中也是如此。我们使用了同基因酵母菌株，其中对P53依赖的反式激活被测量为染色体整合的P53反应元件(RES)。选择10个RE对人P53的DNA结合亲和力和反式激活能力进行了广泛的采样，并利用可诱导表达系统在两个水平上表达了蛋白质。我们表明，该方法能够研究青蛙P53的温度敏感性，并且包含异位反式激活结构域的嵌合结构可以迅速开发出来，以增强蛋白质的活性，例如果蝇P53，这些蛋白质在参与酵母转录机制方面效果不佳。对每种P53蛋白的相对反式激活曲线的变化进行了测量，并与人类P53蛋白的曲线进行了比较。这些结果在很大程度上独立于相对的P53蛋白水平，揭示了P53反式激活特异性的广泛进化差异，甚至在人和鼠的P53之间也是如此。果蝇和人P53在RE中的区分度最大，斑马鱼P53的选择性最低。
Co-evolution of transcription factors (TFs) with their respective cis-regulatory network enhances functional diversity in the course of evolution. We present a new approach to investigate transactivation capacity of sequence-specific TFs in evolutionary studies. Saccharomyces cerevisiae was used as an in vivo test tube and p53 proteins derived from human and five commonly used animal models were chosen as proof of concept. p53 is a highly conserved master regulator of environmental stress responses. Previous reports indicated conserved p53 DNA binding specificity in vitro, even for evolutionary distant species. We used isogenic yeast strains where p53-dependent transactivation was measured towards chromosomally integrated p53 response elements (REs). Ten REs were chosen to sample a wide range of DNA binding affinity and transactivation capacity for human p53 and proteins were expressed at two levels using an inducible expression system. We showed that the assay is amenable to study thermo-sensitivity of frog p53, and that chimeric constructs containing an ectopic transactivation domain could be rapidly developed to enhance the activity of proteins, such as fruit fly p53, that are poorly effective in engaging the yeast transcriptional machinery. Changes in the profile of relative transactivation towards the ten REs were measured for each p53 protein and compared to the profile obtained with human p53. These results, which are largely independent from relative p53 protein levels, revealed widespread evolutionary divergence of p53 transactivation specificity, even between human and mouse p53. Fruit fly and human p53 exhibited the largest discrimination among REs while zebrafish p53 was the least selective.
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影响因子：
64.5
作者：
Deutsch GB;Zielonka EM;Coutandin D;Weber TA;Schäfer B;Hannewald J;Luh LM;Durst FG;Ibrahim M;Hoffmann J;Niesen FH;Sentürk A;Kunkel H;Brutschy B;Schleiff E;Knapp S;Acker-Palmer A;Grez M;McKeon F;Dötsch V
通讯作者：
Dötsch V
影响因子：
3.7
作者：
Brandt, Tobias;Kaar, Joel L.;Veprintsev, Dmitry B.
通讯作者：
Veprintsev, Dmitry B.
影响因子：
2.6
作者：
GIETZ, RD;SCHIESTL, RH;WOODS, RA
通讯作者：
WOODS, RA
影响因子：
14.9
作者：
Beno, Itai;Rosenthal, Karin;Haran, Tali E.
通讯作者：
Haran, Tali E.
影响因子：
3.8
作者：
Bisio A;De Sanctis V;Del Vescovo V;Denti MA;Jegga AG;Inga A;Ciribilli Y
通讯作者：
Ciribilli Y