IKKα和β光遗传聚类激活NF-κB信号。

IF 2.6 3区 生物学 Q3 MATERIALS SCIENCE, BIOMATERIALS
Alexandra Anna Maria Fischer, Markus Michael Kramer, Miguel Baños, Merlin Moritz Grimm, Manfred Fliegauf, Bodo Grimbacher, Gerald Radziwill, Sven Rahmann, Wilfried Weber
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引用次数: 0

摘要

分子光遗传学允许控制响应光的分子信号通路。这使得以空间和时间分辨的方式分析信号激活和传播的动力学成为可能。这种控制的一个关键策略是信号分子的光诱导聚类,这导致它们的激活和随后的下游信号。在这项工作中,开发了一种光遗传学方法来诱导融合到eGFP(一种广泛使用的蛋白质标签)上的不同蛋白质的梯度聚类。为此,一个egfp特异性纳米体被融合到隐花色素2的变体中,以适应不同的簇形成顺序。这可以通过聚集eGFP-IKKα和eGFP-IKKβ来证明,从而实现NF-κB信号的有效和可逆激活。通过RNA测序分析表明,该方法可以通过内源性NF-κB途径激活下游信号传导,从而能够激活NF-κB应答的报告基因结构以及内源性NF-κB应答的靶基因。该系统的通用设计可能可转移到其他信号通路,以分析信号激活和传播的动力学。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Activation of NF-κB Signaling by Optogenetic Clustering of IKKα and β

Activation of NF-κB Signaling by Optogenetic Clustering of IKKα and β

Activation of NF-κB Signaling by Optogenetic Clustering of IKKα and β

Activation of NF-κB Signaling by Optogenetic Clustering of IKKα and β

Activation of NF-κB Signaling by Optogenetic Clustering of IKKα and β

Molecular optogenetics allows the control of molecular signaling pathways in response to light. This enables the analysis of the kinetics of signal activation and propagation in a spatially and temporally resolved manner. A key strategy for such control is the light-inducible clustering of signaling molecules, which leads to their activation and subsequent downstream signaling. In this work, an optogenetic approach is developed for inducing graded clustering of different proteins that are fused to eGFP, a widely used protein tag. To this aim, an eGFP-specific nanobody is fused to Cryptochrome 2 variants engineered for different orders of cluster formation. This is exemplified by clustering eGFP-IKKα and eGFP-IKKβ, thereby achieving potent and reversible activation of NF-κB signaling. It is demonstrated that this approach can activate downstream signaling via the endogenous NF-κB pathway and is thereby capable of activating both an NF-κB-responsive reporter construct as well as endogenous NF-κB-responsive target genes as analyzed by RNA sequencing. The generic design of this system is likely transferable to other signaling pathways to analyze the kinetics of signal activation and propagation.

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来源期刊
Advanced biology
Advanced biology Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)
CiteScore
6.60
自引率
0.00%
发文量
130
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