Optogenetic protein clustering and signaling activation in mammalian cells

@article{Bugaj2013OptogeneticPC,
  title={Optogenetic protein clustering and signaling activation in mammalian cells},
  author={Lukasz J. Bugaj and Atri Choksi and Colin K Mesuda and Ravi S. Kane and David V. Schaffer},
  journal={Nature Methods},
  year={2013},
  volume={10},
  pages={249-252}
}
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… Expand
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References

SHOWING 1-10 OF 38 REFERENCES
Rapid blue light induction of protein interactions in living cells
TLDR
Genetically encoded light-inducible protein-interaction modules based on Arabidopsis thaliana cryptochrome 2 and CIB1 that require no exogenous ligands and dimerize on blue-light exposure with subsecond time resolution and subcellular spatial resolution are described. Expand
TULIPs: Tunable, light-controlled interacting protein tags for cell biology
TLDR
T tunable light-inducible dimerization tags (TULIPs) based on a synthetic interaction between the LOV2 domain of Avena sativa phototropin 1 and an engineered PDZ domain (ePDZ) are developed. Expand
Spatiotemporal Control of Cell Signalling Using A Light-Switchable Protein Interaction
TLDR
It is shown that light-gated translocation of the upstream activators of Rho-family GTPases, which control the actin cytoskeleton, can be used to precisely reshape and direct the cell morphology of mammalian cells. Expand
Induction of protein-protein interactions in live cells using light
TLDR
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. Expand
Engineered photoreceptors as novel optogenetic tools.
  • Andreas Möglich, K. Moffat
  • Biology, Medicine
  • Photochemical & photobiological sciences : Official journal of the European Photochemistry Association and the European Society for Photobiology
  • 2010
TLDR
Basic signaling principles are treated and the two photosensor classes which are currently most widely used in fusion-based design: LOV domains and phytochromes are discussed and general strategies for the engineering of photoreceptors are developed. Expand
Rho GTPases: biochemistry and biology.
  • A. Jaffe, A. Hall
  • Biology, Medicine
  • Annual review of cell and developmental biology
  • 2005
TLDR
This review presents the best characterized of these biochemical pathways that control some of the most fundamental processes of cell biology common to all eukaryotes, including morphogenesis, polarity, movement, and cell division. Expand
A genetically-encoded photoactivatable Rac controls the motility of living cells
TLDR
A new approach to produce genetically encoded photoactivatable derivatives of Rac1, a key GTPase regulating actin cytoskeletal dynamics in metazoan cells, which was shown to inhibit RhoA in mouse embryonic fibroblasts, with inhibition modulated at protrusions and ruffles. Expand
Functional interaction of phytochrome B and cryptochrome 2
TLDR
The genetic interaction that occurs between the Arabidopsis photoreceptors phyB and cry2 is shown in the control of flowering time, hypocotyl elongation and circadian period by the clock and fluorescent resonance energy transfer microscopy is used to show that phyBs interact in nuclear speckles that are formed in a light-dependent fashion. Expand
Photoexcited CRY2 Interacts with CIB1 to Regulate Transcription and Floral Initiation in Arabidopsis
TLDR
The identification and characterization of the Arabidopsis CIB1 (cryptochrome-interacting basic-helix-loop-Helix) protein is reported, and it is proposed that the blue light–dependent interaction of cryptochrome(s) with C IB1 and CIB 1-related proteins represents an early photoreceptor signaling mechanism in plants. Expand
Controlling signal transduction with synthetic ligands.
TLDR
Cell permeable, synthetic ligands were devised that can be used to control the intracellular oligomerization of specific proteins to demonstrate their utility and have the potential to be applied wherever precise control of a signal transduction pathway is desired. Expand
...
1
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3
4
...