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Spatio‐temporally precise activation of engineered receptor tyrosine kinases by light

Michael Grusch, Karin Schelch, Robert Riedler, Eva Reichhart, Christopher Differ, Walter Berger, Álvaro Inglés‐Prieto, Harald Janovjak

Author Affiliations

  1. Michael Grusch1,,
  2. Karin Schelch1,,
  3. Robert Riedler2,,
  4. Eva Reichhart2,
  5. Christopher Differ2,
  6. Walter Berger1,
  7. Álvaro Inglés‐Prieto2 and
  8. Harald Janovjak*,2
  1. 1Department of Medicine I, Institute of Cancer Research, Comprehensive Cancer Center Vienna, Medical University of Vienna, Vienna, Austria
  2. 2Institute of Science and Technology Austria (IST Austria), Klosterneuburg, Austria
  1. *Corresponding author. Tel: +43 2243 9000 4201; Fax: +43 2243 9000 2000; E‐mail: harald{at}ist.ac.at
  1. These authors contributed equally and share first authorship

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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.

Synopsis

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A new method for spatially and temporally controlled activation of receptor tyrosine kinases (RTKs) is described. Bacterial LOV domains allow blue light‐activated dimerization of ligand‐insensitive FGFR, EGFR or RET receptors and mitogenic signalling.

  • Light‐oxygen‐voltage (LOV)‐sensing domains induce protein dimerization when activated by light.

  • Engineered FGFR1, EGFR and RET receptor tyrosine kinases that incorporate LOV domains self‐dimerize and get activated upon light exposure.

  • Light‐mediated activation of RTKs induces the respective endogenous downstream signalling pathways with high temporal and spatial resolution.

  • Light‐mediated activation of RTKs mimics complex cellular behaviours normally regulated by the physiological ligands of the RTKs.

The EMBO Journal (2014) 33: 1713–1726

  • Received December 15, 2013.
  • Revision received May 22, 2014.
  • Accepted May 26, 2014.
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