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Induction of protein-protein interactions in live cells using light

Nature Biotechnology volume 27, pages 941–945 (2009)Cite this article

Abstract

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.

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Figure 1: GI and FKF1 interact in response to blue light.
Figure 2: Light-induced recruitment of FKF1-Rac1 to the plasma membrane and formation of lamellipodia.
Figure 3: Light-induced activation of transcription.

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Acknowledgements

We would like to thank T. Imaizumi (Scripps Research Institute) for providing FKF1 and GI plasmids; T.-S. Tseng and W.R. Briggs (Carnegie Institution, Stanford University) for providing phototropin plasmids; Y. Gotoh (University of Tokyo) for providing plasmids encoding human Rac1 mutants; and M. Endo and S. Sugano for helpful advice. This study is supported by an National Institutes of Health Pioneer Award, the Simons Foundation and by gifts from L. Miller, B. and F. Horowitz and M. MCafferey to R.E.D., the Japan Society for the Promotion of Science Postdoctoral Fellowships for Research Abroad to M.Y. and Stanford Institutes of Medicine Summer Research Program to B.H.

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  1. Brian Hsueh

    Present address: Present address: Department of Chemistry, Princeton University, Princeton, New Jersey, USA.,

Authors and Affiliations

  1. Department of Neurobiology, Stanford University School of Medicine, Stanford, California, USA

    Masayuki Yazawa, Amir M Sadaghiani, Brian Hsueh & Ricardo E Dolmetsch

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  1. Masayuki Yazawa
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  2. Amir M Sadaghiani
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Contributions

M.Y. and R.E.D. designed research and wrote manuscript; M.Y. performed plasmid construction, live-cell imaging and data analysis; M.Y. and A.M.S. conducted mutagenesis of FKF1; M.Y. and B.H. performed immunocytochemistry and luciferase assays.

Corresponding author

Correspondence to Ricardo E Dolmetsch.

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Yazawa, M., Sadaghiani, A., Hsueh, B. et al. Induction of protein-protein interactions in live cells using light. Nat Biotechnol 27, 941–945 (2009). https://doi.org/10.1038/nbt.1569

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