Optogenetic Control Reveals Differential Promoter Interpretation of Transcription Factor Nuclear Translocation Dynamics.
Optogenetic Control Reveals Differential Promoter Interpretation of Transcription Factor Nuclear Translocation Dynamics.
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DOI：
10.1016/j.cels.2020.08.009
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发表时间：
2020-10-21
期刊：
影响因子：
9.3
通讯作者：
El-Samad H
中科院分区：
文献类型：
--
作者：
Chen SY;Osimiri LC;Chevalier M;Bugaj LJ;Nguyen TH;Greenstein RA;Ng AH;Stewart-Ornstein J;Neves LT;El-Samad H
基因表达被认为不仅受转录因子（TF）浓度的影响，而且受其核转位动力学的影响。测试这个假设需要直接控制TF动态。在这里，我们设计CLASP，这是一种用于快速和可调的感兴趣TF易位的光遗传学工具。使用CLASP融合到Crz1，我们观察到，随着时间的推移，对于相同的集成浓度的核TF，改变输入动态改变靶基因表达：脉冲输入产生更高的表达比连续输入，反之亦然，这取决于靶基因。计算模型表明，在低TF输入饱和的剂量响应可以产生更高的基因表达的脉冲与连续输入，和多态启动子激活可以产生相反的行为。我们的集成工具开发和建模方法表征了Crz1核转位动力学的启动子响应，提取了可能有助于解释靶基因差异表达的定量特征。CLASP是控制转录因子（TF）的核定位并从其同源启动子引发基因表达的模块化光遗传学策略。CLASP控制的Crz1核定位，再加上计算建模，揭示了启动子如何可以差异解码动态转录因子信号。CLASP开发和建模的综合策略提供了一种通用的方法来因果调查动态TF核穿梭的转录后果。
Gene expression is thought to be affected not only by the concentration of transcription factors (TFs) but also the dynamics of their nuclear translocation. Testing this hypothesis requires direct control of TF dynamics. Here, we engineer CLASP, an optogenetic tool for rapid and tunable translocation of a TF of interest. Using CLASP fused to Crz1, we observe that, for the same integrated concentration of nuclear TF over time, changing input dynamics changes target gene expression: pulsatile inputs yield higher expression than continuous inputs, or vice versa, depending on the target gene. Computational modeling reveals that a dose-response saturating at low TF input can yield higher gene expression for pulsatile versus continuous input, and that multi-state promoter activation can yield the opposite behavior. Our integrated tool development and modeling approach characterize promoter responses to Crz1 nuclear translocation dynamics, extracting quantitative features that may help explain the differential expression of target genes. CLASP is a modular optogenetic strategy to control the nuclear localization of transcription factors (TFs) and elicit gene expression from their cognate promoters. CLASP control of Crz1 nuclear localization, coupled with computational modeling, revealed how promoters can differentially decode dynamic transcription factor signals. The integrated strategy of CLASP development and modeling presents a generalized approach to causally investigate the transcriptional consequences of dynamic TF nuclear shuttling.
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