Optogenetic protein clustering and signaling activation in mammalian cells

Nat Methods. 2013 Mar;10(3):249-52. doi: 10.1038/nmeth.2360. Epub 2013 Feb 3.

Abstract

We report an optogenetic method based on Arabidopsis thaliana cryptochrome 2 for rapid and reversible protein oligomerization in response to blue light. We demonstrated its utility by photoactivating the β-catenin pathway, achieving a transcriptional response higher than that obtained with the natural ligand Wnt3a. We also demonstrated the modularity of this approach by photoactivating RhoA with high spatiotemporal resolution, thereby suggesting a previously unknown mode of activation for this Rho GTPase.

Publication types

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

MeSH terms

  • Animals
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Arabidopsis Proteins / radiation effects
  • Blotting, Western
  • Cell Culture Techniques
  • Cell Nucleus / metabolism
  • Cell Nucleus / radiation effects
  • Cryptochromes / genetics
  • Cryptochromes / metabolism*
  • Cryptochromes / radiation effects
  • Cytoplasm / metabolism
  • Cytoplasm / radiation effects
  • Enzyme-Linked Immunosorbent Assay
  • Fluorescence Recovery After Photobleaching
  • HEK293 Cells
  • Humans
  • Light
  • Light Signal Transduction
  • Luminescent Proteins / metabolism
  • Luminescent Proteins / radiation effects
  • Mice
  • NIH 3T3 Cells
  • Protein Multimerization* / radiation effects
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Fusion Proteins / radiation effects
  • Transcription, Genetic
  • Transfection
  • Wnt Signaling Pathway* / radiation effects
  • Wnt3A Protein / genetics
  • Wnt3A Protein / metabolism*
  • Wnt3A Protein / radiation effects
  • beta Catenin / genetics
  • beta Catenin / metabolism*
  • beta Catenin / radiation effects
  • rhoA GTP-Binding Protein / genetics
  • rhoA GTP-Binding Protein / metabolism*
  • rhoA GTP-Binding Protein / radiation effects

Substances

  • Arabidopsis Proteins
  • CRY2 protein, Arabidopsis
  • Cryptochromes
  • Luminescent Proteins
  • Recombinant Fusion Proteins
  • Wnt3A Protein
  • beta Catenin
  • red fluorescent protein
  • rhoA GTP-Binding Protein