Tunable transcription factor library for robust quantification of regulatory properties in Escherichia coli.
Tunable transcription factor library for robust quantification of regulatory properties in Escherichia coli.
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DOI：
10.15252/msb.202110843
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发表时间：
2022-06
影响因子：
9.9
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基于结合序列、结合位置和启动子类型等因素预测转录因子的定量调控功能是不可能的。基因网络的互连性质和调节单个转铁蛋白浓度的困难使得对转铁蛋白功能的孤立研究具有挑战性。在这里，我们提出了一个大肠杆菌菌株的文库，旨在允许精确控制单个TF的浓度，从而能够研究TF浓度对生理和调节的作用。我们通过测量锌响应的Tf，ZntR和平行的Tf对，GalR/GALS的调节功能，证明了这一资源的有效性。对于锌受体，我们发现锌以一种方式改变了锌受体的调节功能，使得被调控基因的激活相对于锌受体浓度是健壮的。对于GalR和GALS，我们能够证明这些平行的TF除了结合亲和力方面的差异外，还具有根本上不同的调节作用。本研究提供了一个大肠杆菌菌株文库，其中单个转录因子(TF)的浓度可以独立调节和测量，从而能够分析依赖于拷贝数的TF在调控网络和细胞生理学中的作用。
Predicting the quantitative regulatory function of transcription factors (TFs) based on factors such as binding sequence, binding location, and promoter type is not possible. The interconnected nature of gene networks and the difficulty in tuning individual TF concentrations make the isolated study of TF function challenging. Here, we present a library of Escherichia coli strains designed to allow for precise control of the concentration of individual TFs enabling the study of the role of TF concentration on physiology and regulation. We demonstrate the usefulness of this resource by measuring the regulatory function of the zinc‐responsive TF, ZntR, and the paralogous TF pair, GalR/GalS. For ZntR, we find that zinc alters ZntR regulatory function in a way that enables activation of the regulated gene to be robust with respect to ZntR concentration. For GalR and GalS, we are able to demonstrate that these paralogous TFs have fundamentally distinct regulatory roles beyond differences in binding affinity. This study presents a library of Escherichia coli strains where the concentration of individual transcription factors (TFs) can be independently tuned and measured, allowing for the analysis of copy number‐dependent TF roles in regulatory networks and cellular physiology.