A High-Throughput Mutational Scan of an Intrinsically Disordered Acidic Transcriptional Activation Domain.
A High-Throughput Mutational Scan of an Intrinsically Disordered Acidic Transcriptional Activation Domain.
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DOI：
10.1016/j.cels.2018.01.015
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发表时间：
2018-04-25
期刊：
影响因子：
9.3
通讯作者：
Cohen BA
中科院分区：
文献类型：
--
作者：
Staller MV;Holehouse AS;Swain-Lenz D;Das RK;Pappu RV;Cohen BA
转录活化结构域对基因调控至关重要，但其内在的无序性和低一级序列保守性使得鉴定其活性背后的氨基酸组成特征变得困难。在这里，我们描述了一种合理的诱变方案，该方案通过量化体内数千种变体的活性并使用全原子蒙特卡罗方法模拟它们的构象集合来反卷积四个激活域序列特征（酸性、疏水性、内在无序和短线性基序）的功能。我们的研究结果与酿酒酵母转录因子Gcn4的典型激活域相一致，将现有的观察结果统一为其功能的统一模型：内在无序和酸性残基使两个疏水基序免于驱动崩溃。相反，最活跃的变体将其芳香残留物暴露在溶剂中。我们的研究结果说明了如何通过高通量的合理诱变揭示内在无序蛋白的功能。自从30多年前被发现以来，人们一直不清楚为什么许多转录因子的转录激活域是酸性的。Staller等人描述了一种方法来测量数千个合理设计的激活结构域突变体的活性。用这种方法，他们揭示了经典模型激活结构域的酸性残基的作用。
Transcriptional activation domains are essential for gene regulation but their intrinsic disorder and low primary sequence conservation have made it difficult to identify the amino acid composition features that underlie their activity. Here, we describe a rational mutagenesis scheme that deconvolves the function of four activation domain sequence features — acidity, hydrophobicity, intrinsic disorder, and short linear motifs — by quantifying the activity of thousands of variants in vivo and simulating their conformational ensembles using an all-atom Monte Carlo approach. Our results with a canonical activation domain from the Saccharomyces cerevisiae transcription factor Gcn4, reconcile existing observations into a unified model of its function: the intrinsic disorder and acidic residues keep two hydrophobic motifs from driving collapse. Instead, the most active variants keep their aromatic residues exposed to the solvent. Our results illustrate how the function of intrinsically disordered proteins can be revealed by high-throughput rational mutagenesis. Since their discovery over 30 years ago, it has been unclear why many transcriptional activation domains of transcription factors are acidic. Staller et al. describe a method to measure the activities of thousands of rationally designed activation domains mutants‥ With this method they uncover a role for the acidic residues of a classic model activation domain.
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影响因子：
48
作者：
Fowler, Douglas M.;Fields, Stanley
通讯作者：
Fields, Stanley
DOI：
10.1038/nrg2847
发表时间：
2010-08
期刊：
Nature reviews. Genetics
影响因子：
--
作者：
通讯作者：
--
影响因子：
8
作者：
Frank, BS;Vardar, D;McKnight, CJ
通讯作者：
McKnight, CJ
影响因子：
3.5
作者：
Cormack, BP;Valdivia, RH;Falkow, S
通讯作者：
Falkow, S
影响因子：
14.8
作者：
Borcherds, Wade;Theillet, Francois-Xavier;Daughdrill, Gary W.
通讯作者：
Daughdrill, Gary W.