Spatiotemporal Control of Cell Signalling Using A Light-Switchable Protein Interaction

By Anselm Levskaya, O. Weiner, Wendell A. Lim, Christopher A. Voigt. Nature, 2009

998 citations recorded by the source corpus.

Genetically encodable optical reporters, such as green fluorescent protein, have revolutionized the observation and measurement of cellular states. However, the inverse challenge of using light to control precisely cellular behaviour has only recently begun to be addressed; semi-synthetic chromophore-tethered receptors and naturally occurring channel rhodopsins have been used to perturb directly neuronal networks. The difficulty of engineering light-sensitive proteins remains a significant impediment to the optical control of most cell-biological processes. Here we demonstrate the use of a new genetically encoded light-control system based on an optimized, reversible protein–protein interaction from the phytochrome signalling network of Arabidopsis thaliana. Because protein–protein interactions are one of the most general currencies of cellular information, this system can, in principle, be generically used to control diverse functions. Here we show that this system can be used to translocate target proteins precisely and reversibly to the membrane with micrometre spatial resolution and at the second timescale. We show 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. The light-gated protein–protein interaction that has been optimized here should be useful for the design of diverse light-programmable reagents, potentially enabling a new generation of perturbative, quantitative experiments in cell biology.

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Citations in the corpus

Engineering an improved light-induced dimer (iLID) for controlling the localization and activity of signaling proteins (2014, background): “PhyB and PIF interact after irradiation with red light and dissociate with exposure to far-red light (3).”
Encoding and Decoding Cellular Information through Signaling Dynamics (2013, unknown): “There has been recent work demonstrating light-based perturbations to cellular 954 Cell 152, February 28, 2013 ª2013 Elsevier Inc. dynamics (Levskaya et al., 2009; Toettcher et al., 2011), which could provide exceptionally noninvasive and precise control over temporal signaling.”
Molecular and cellular approaches for diversifying and extending optogenetics. (2010, background): “…with high dark activity that limits in vivo application (Schröder-Lang et al., 2007), and subsequent work on light-sensitive PAS or LOV domains (Levskaya et al., 2009; Wu et al., 2009) may open up new ways to control protein-protein association if these approaches can be made to operate in…”
Synthetic biology: applications come of age (2010, methodology): “Using a synthetic hybrid approach, Voigt and colleague”
The Development and Application of Optogenetics (2011, background): “Optical control over small GTPases was next achieved in cultured cells by several different laboratories (Levskaya et al. 2009, Wu et al. 2009, Yazawa et al. 2009) using optically modulated protein-protein interactions; although these have not yet been shown to express or display single-component…”
Spatiotemporal control of intracellular phase transitions using light-activated optoDroplets (2016, unknown): “The field has primarily focused on precise control over homo-or hetero-dimerization ((Toettcher et al., 2011), (Kennedy et al., 2010), (Levskaya et al., 2009)).”
CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes (2013, unknown): “For example, dCas9 could be used with existing optogenetic or chemicalinduced proximity systems to dynamically analyze recruitment of a broad range of chromatin modifiers to a specific DNA sequence (Hathaway et al., 2012; Levskaya et al., 2009).”
Development of an optimized backbone of FRET biosensors for kinases and GTPases (2011, background): “In a preliminary experiment, linkers consisting of (SAGG)n, where n indicates the number of repeats, served better than the 72 a.a. polyglycine linkers (Levskaya et al., 2009; data not shown).”