Spatio-temporally precise activation of engineered receptor tyrosine kinases by light Journal Article

Author(s): Grusch, Michael; Schelch, Karin; Riedler, Robert; Reichhart, Eva; Differ, Christopher; Berger, Walter; Ingles-Prieto, Álvaro; Janovjak, Harald
Article Title: Spatio-temporally precise activation of engineered receptor tyrosine kinases by light
Affiliation IST Austria
Abstract: Receptor tyrosine kinases (RTKs) are a large family of cell surface receptors that sense growth factors and hormones and regulate a variety of cell behaviours in health and disease. Contactless activation of RTKs with spatial and temporal precision is currently not feasible. Here, we generated RTKs that are insensitive to endogenous ligands but can be selectively activated by low-intensity blue light. We screened light-oxygen-voltage (LOV)-sensing domains for their ability to activate RTKs by light-activated dimerization. Incorporation of LOV domains found in aureochrome photoreceptors of stramenopiles resulted in robust activation of the fibroblast growth factor receptor 1 (FGFR1), epidermal growth factor receptor (EGFR) and rearranged during transfection (RET). In human cancer and endothelial cells, light induced cellular signalling with spatial and temporal precision. Furthermore, light faithfully mimicked complex mitogenic and morphogenic cell behaviour induced by growth factors. RTKs under optical control (Opto-RTKs) provide a powerful optogenetic approach to actuate cellular signals and manipulate cell behaviour.
Keywords: Optogenetics; aureochrome; cell signalling; light-oxygen-voltage (LOV)-sensing domain; synthetic biology; synthetic physiology
Journal Title: EMBO Journal
Volume: 33
Issue 15
ISSN: 1460-2075
Publisher: Wiley-Blackwell  
Date Published: 2014-08-01
Start Page: 1713
End Page: 1726
Sponsor: European Union Seventh Framework Programme; Human Frontier Science Program; Oesterreichische Nationalbank Anniversary Fund 14211; Austrian Research Promotion Agency; FemTech
DOI: 10.15252/embj.201387695
Notes: We thank G. Krupitza, C. McKenzie, D. Siekhaus and M. Matveenko for discussions, E. Papusheva for help with microscopy, M. Spanova, A Kitzmann, B. Peter‐Vörösmarty and D. Meindl for technical assistance, V. Baleyeava and S. zur Nedden for assistance with genetic engineering, Ariad Pharmaceuticals Inc. for AP20187, D.M. Spencer (Baylor College of Medicine) and Y. Yang (East China University of Science and Technology) for plasmids, V.L. Kinnula (University of Helsinki) and R. Stahel (University of Zurich) for mesothelioma cells, and S. Geleff (Medical University of Vienna) for hBE cells.
Open access: yes (repository)