An optogenetic system for interrogating the temporal dynamics of Akt.
An optogenetic system for interrogating the temporal dynamics of Akt.
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DOI：
10.1038/srep14589
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发表时间：
2015-10-01
影响因子：
4.6
通讯作者：
Ozawa T
中科院分区：
文献类型：
--
作者：
Katsura Y;Kubota H;Kunida K;Kanno A;Kuroda S;Ozawa T
丝氨酸/苏氨酸激酶Akt的动态活性对于调节多种细胞功能至关重要，但其活性的精确时空控制仍然是一个关键问题。本研究基于拟南芥隐花色素2（cryptochrome 2，CIB 1）的光诱导蛋白相互作用模块，构建了一个光激活Akt（PA-Akt）系统。Akt融合到Akt 2 phr，其是Akt 2的最小光敏结构域（Akt 2-Akt），在生理动态范围内通过光照在几分钟内可逆地激活，并且特异性地调节下游分子和诱导的生物学功能。我们已经产生了一个计算模型的PA 2-Akt激活，使我们能够使用PA-Akt来控制活动定量。该系统提供的证据表明，Akt活性的时间模式对于产生Akt-FoxO途径的下游功能之一至关重要;参与肌肉萎缩的关键基因（Atrogin-1）的表达。光学模块与计算建模的使用代表了通过光遗传学模块的预测操作来询问生物分子的时间动态的一般框架。
The dynamic activity of the serine/threonine kinase Akt is crucial for the regulation of diverse cellular functions, but the precise spatiotemporal control of its activity remains a critical issue. Herein, we present a photo-activatable Akt (PA-Akt) system based on a light-inducible protein interaction module of Arabidopsis thaliana cryptochrome2 (CRY2) and CIB1. Akt fused to CRY2phr, which is a minimal light sensitive domain of CRY2 (CRY2-Akt), is reversibly activated by light illumination in several minutes within a physiological dynamic range and specifically regulates downstream molecules and inducible biological functions. We have generated a computational model of CRY2-Akt activation that allows us to use PA-Akt to control the activity quantitatively. The system provides evidence that the temporal patterns of Akt activity are crucial for generating one of the downstream functions of the Akt-FoxO pathway; the expression of a key gene involved in muscle atrophy (Atrogin-1). The use of an optical module with computational modeling represents a general framework for interrogating the temporal dynamics of biomolecules by predictive manipulation of optogenetic modules.