Red Light-Regulated Reversible Nuclear Localization of Proteins in Mammalian Cells and Zebrafish.

Protein trafficking in and out of the nucleus represents a key step in controlling cell fate and function. Here we report the development of a red light-inducible and far-red light-reversible synthetic system for controlling nuclear localization of proteins in mammalian cells and zebrafish. First, we synthetically reconstructed and validated the red light-dependent Arabidopsis phytochrome B nuclear import mediated by phytochrome-interacting factor 3 in a nonplant environment and support current hypotheses on the import mechanism in planta. On the basis of this principle we next regulated nuclear import and activity of target proteins by the spatiotemporal projection of light patterns. A synthetic transcription factor was translocated into the nucleus of mammalian cells and zebrafish to drive transgene expression. These data demonstrate the first in vivo application of a plant phytochrome-based optogenetic tool in vertebrates and expand the repertoire of available light-regulated molecular devices.

[1]  A. Deiters,et al.  Control of Protein Function through Optochemical Translocation , 2014, ACS synthetic biology.

[2]  Matias D. Zurbriggen,et al.  Orthogonal optogenetic triple-gene control in Mammalian cells. , 2014, ACS synthetic biology.

[3]  Jens Timmer,et al.  Multi-chromatic control of mammalian gene expression and signaling , 2013, Nucleic acids research.

[4]  M. Kennedy,et al.  A light-triggered protein secretion system , 2013, The Journal of cell biology.

[5]  S. Guan,et al.  Multisite Light-Induced Phosphorylation of the Transcription Factor PIF3 Is Necessary for Both Its Rapid Degradation and Concomitant Negative Feedback Modulation of Photoreceptor phyB Levels in Arabidopsis[C][W] , 2013, Plant Cell.

[6]  E. Schäfer,et al.  Interaction with plant transcription factors can mediate nuclear import of phytochrome B , 2012, Proceedings of the National Academy of Sciences.

[7]  Michael Z. Lin,et al.  Optical Control of Protein Activity by Fluorescent Protein Domains , 2012, Science.

[8]  B. Kuhlman,et al.  A genetically-encoded photoactivatable Rac controls the motility of living cells , 2009, Nature.

[9]  J. Casal.  Photoreceptor signaling networks in plant responses to shade. , 2013, Annual review of plant biology.

[10]  Ravi S Kane,et al.  Optogenetic protein clustering and signaling activation in mammalian cells , 2013, Nature Methods.

[11]  Matias D Zurbriggen,et al.  Optogenetics for gene expression in mammalian cells , 2015, Biological chemistry.

[12]  Matias D Zurbriggen,et al.  Control of gene expression using a red- and far-red light–responsive bi-stable toggle switch , 2014, Nature Protocols.

[13]  Jens Timmer,et al.  A red/far-red light-responsive bi-stable toggle switch to control gene expression in mammalian cells , 2013, Nucleic acids research.

[14]  Christopher A. Voigt,et al.  Spatiotemporal Control of Cell Signalling Using A Light-Switchable Protein Interaction , 2009, Nature.

[15]  Matias D Zurbriggen,et al.  Optogenetic control of protein kinase activity in mammalian cells. , 2014, ACS synthetic biology.

[16]  J. Dinman,et al.  Systematic analysis of bicistronic reporter assay data , 2004, Nucleic acids research.

[17]  M. Ehlers,et al.  Rapid blue light induction of protein interactions in living cells , 2010, Nature Methods.

[18]  Andreas Möglich,et al.  Engineering of a red-light–activated human cAMP/cGMP-specific phosphodiesterase , 2014, Proceedings of the National Academy of Sciences.

[19]  Yi Yang,et al.  Spatiotemporal control of gene expression by a light-switchable transgene system , 2012, Nature Methods.

[20]  Hannes M. Beyer,et al.  Optogenetic control of signaling in mammalian cells , 2015, Biotechnology journal.

[21]  Orion D. Weiner,et al.  Illuminating cell signalling with optogenetic tools , 2014, Nature Reviews Molecular Cell Biology.

[22]  J. Epstein,et al.  Signal transduction: An eye on organ development , 2003, Nature.

[23]  Martin Fussenegger,et al.  Mind-controlled transgene expression by a wireless-powered optogenetic designer cell implant , 2014, Nature Communications.

[24]  Ekaterina A. Kostina,et al.  Photo-sensitive degron variants for tuning protein stability by light , 2014, BMC Systems Biology.

[25]  Shuji Ogino,et al.  Restriction of intestinal stem cell expansion and the regenerative response by YAP , 2012, Nature.

[26]  Chao Tang,et al.  A light-inducible organelle-targeting system for dynamically activating and inactivating signaling in budding yeast , 2013, Molecular biology of the cell.

[27]  Mark Gomelsky,et al.  Engineering adenylate cyclases regulated by near-infrared window light , 2014, Proceedings of the National Academy of Sciences.

[28]  M. Neuberger,et al.  The stability of AID and its function in class-switching are critically sensitive to the identity of its nuclear-export sequence , 2009, Proceedings of the National Academy of Sciences.

[29]  Young Hun Song,et al.  LOV domain-containing F-box proteins: light-dependent protein degradation modules in Arabidopsis. , 2012, Molecular plant.

[30]  Eberhard Schäfer,et al.  Perception of UV-B by the Arabidopsis UVR8 Protein , 2011, Science.

[31]  R. Eils,et al.  Engineering light-inducible nuclear localization signals for precise spatiotemporal control of protein dynamics in living cells , 2014, Nature Communications.

[32]  L. Regan,et al.  Antiparallel Leucine Zipper-Directed Protein Reassembly: Application to the Green Fluorescent Protein , 2000 .

[33]  Matias D. Zurbriggen,et al.  Synthesis of phycocyanobilin in mammalian cells. , 2013, Chemical communications.

[34]  J. Chory,et al.  Regulation of Phytochrome B Nuclear Localization through Light-Dependent Unmasking of Nuclear-Localization Signals , 2005, Current Biology.

[35]  K. Aldape,et al.  Nuclear PKM2 regulates β-catenin transactivation upon EGFR activation , 2011, Nature.

[36]  Pamela A. Silver,et al.  Nuclear transport and cancer: from mechanism to intervention , 2004, Nature Reviews Cancer.

[37]  T. Halazonetis,et al.  Ultraviolet-B-mediated induction of protein–protein interactions in mammalian cells , 2013, Nature Communications.