Regulation of endogenous transmembrane receptors through optogenetic Cry2 clustering

Nat Commun. 2015 Apr 22:6:6898. doi: 10.1038/ncomms7898.

Abstract

Transmembrane receptors are the predominant conduit through which cells sense and transduce extracellular information into intracellular biochemical signals. Current methods to control and study receptor function, however, suffer from poor resolution in space and time and often employ receptor overexpression, which can introduce experimental artefacts. We report a genetically encoded approach, termed Clustering Indirectly using Cryptochrome 2 (CLICR), for spatiotemporal control over endogenous transmembrane receptor activation, enabled through the optical regulation of target receptor clustering and downstream signalling using noncovalent interactions with engineered Arabidopsis Cryptochrome 2 (Cry2). CLICR offers a modular platform to enable photocontrol of the clustering of diverse transmembrane receptors including fibroblast growth factor receptor (FGFR), platelet-derived growth factor receptor (PDGFR) and integrins in multiple cell types including neural stem cells. Furthermore, light-inducible manipulation of endogenous receptor tyrosine kinase (RTK) activity can modulate cell polarity and establish phototaxis in fibroblasts. The resulting spatiotemporal control over cellular signalling represents a powerful new optogenetic framework for investigating and controlling cell function and fate.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • 3T3 Cells
  • Animals
  • Arabidopsis Proteins / metabolism*
  • Blotting, Western
  • Cell Membrane / metabolism*
  • Cell Polarity
  • Cryptochromes / metabolism*
  • Fibroblasts
  • HEK293 Cells
  • Humans
  • Immunoprecipitation
  • Light
  • Mice
  • Microscopy, Confocal
  • Optogenetics / methods*
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Signal Transduction

Substances

  • Arabidopsis Proteins
  • CRY2 protein, Arabidopsis
  • Cryptochromes
  • Receptor Protein-Tyrosine Kinases