Induction of protein-protein interactions in live cells using light
Induction of protein-protein interactions in live cells using light
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DOI：
10.1038/nbt.1569
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发表时间：
2009-10-01
影响因子：
46.9
通讯作者：
Dolmetsch, Ricardo E.
中科院分区：
文献类型：
--
作者：
Yazawa, Masayuki;Sadaghiani, Amir M.;Dolmetsch, Ricardo E.
蛋白质-蛋白质相互作用对于许多细胞过程是必不可少的。我们开发了一种称为光激活二聚化（LAD）的技术，利用光在活细胞中人工诱导蛋白质异源和同源二聚化。使用FKF 1和GIGANTEA（GI）蛋白质的拟南芥，我们已经产生的蛋白质标签，其相互作用是由蓝光控制。我们证明了该系统的效用与LAD结构，可以招募小G-蛋白Rac 1的质膜和诱导局部形成的片状伪足响应于焦点照明。我们还通过将GI和FKF 1的结构域分别与Gal 4的DNA结合结构域和VP 16的反式激活结构域融合来产生光激活转录因子，这表明该技术很容易适用于其他系统。这些研究为开发用于控制生物体中受体激活、突触形成和其他信号事件的光调节信号分子奠定了基础。
Protein-protein interactions are essential for many cellular processes. We have developed a technology called light-activated dimerization (LAD) to artificially induce protein hetero- and homodimerization in live cells using light. Using the FKF1 and GIGANTEA (GI) proteins of Arabidopsis thaliana, we have generated protein tags whose interaction is controlled by blue light. We demonstrated the utility of this system with LAD constructs that can recruit the small G-protein Rac1 to the plasma membrane and induce the local formation of lamellipodia in response to focal illumination. We also generated a light-activated transcription factor by fusing domains of GI and FKF1 to the DNA binding domain of Gal4 and the transactivation domain of VP16, respectively, showing that this technology is easily adapted to other systems. These studies set the stage for the development of light-regulated signaling molecules for controlling receptor activation, synapse formation and other signaling events in organisms.