Activation of protein splicing with light in yeast

Abstract

Spatiotemporal regulation of protein function is a key feature of living systems; experimental tools that provide such control are of great utility. Here we report a genetically encoded system for controlling a post-translational process, protein splicing, with light. Studies in Saccharomyces cerevisiae demonstrate that fusion of a photodimerization system from Arabidopsis thaliana to an artificially split intein permits rapid activation of protein splicing to yield a new protein product.

Algal photoreceptors: in vivo functions and potential applications

“…Fusion of an Arabidopsis cryptochrome with a DNA damage checkpoint protein even enabled the activation of the DNA damage signaling pathway in the absence of DNA damage (Ozkan-Dagliyan et al 2013). Also the light-inducible interactions between phytochrome and the phytochrome interaction factor 3 (PIF3) (Leung et al 2008;Tyszkiewicz and Muir 2008) or the LOV-containing F-box proteins FKF1 and its partner GIGANTEA (Yazawa et al 2009) have been used to regulate various cellular activities. A light-switchable gene promoter system should also be mentioned, which was constructed using an Arabidopsis phytochrome; this system allows for reversible control of gene expression with red and far-red light (Shimizu-Sato et al 2002).…”

Engineered pairs of distinct photoswitches for optogenetic control of cellular proteins

“…S everal natural photoreceptors derived from plants, fungi and bacteria, such as the LOV2 domain of phototropin 1, phytochrome B, cryptochrome 2 (CRY2), FKF1, Vivid (VVD), UVR8 and EL222, have been used as photoswitches to construct the optogenetic tool known as a photoactivatable actuator [1][2][3][4][5][6][7][8][9][10] . These photoreceptor-based photoswitches proved that the methods used to control protein activities with light provide a powerful approach to perturbing cellular signalling processes, such as cell morphology [4][5][6] , signal transduction 5 and gene expression 2,3,[6][7][8][9][10] .…”

“…These photoreceptor-based photoswitches proved that the methods used to control protein activities with light provide a powerful approach to perturbing cellular signalling processes, such as cell morphology [4][5][6] , signal transduction 5 and gene expression 2,3,[6][7][8][9][10] . In addition to the photoswitches developed from natural photoreceptors, engineered photoswitches (TULIPs, LOV-ipaA and LOV-SsrA) based on peptide-tagged LOV2 domains and peptide-binding domains/ proteins have been recently developed 11,12 .…”

Spatiotemporally controlled initiation of Parkin-mediated mitophagy within single cells

“…Precise control and perturbation over cellular fate and activities such as neuronal firing, 19,20 activation of specific signaling cascades, [21][22][23] gene transcription, 24 protein production, 25 and modulation of proteinprotein interactions 26 in living cells and organisms have been achieved using light signals. These light-based methods can modulate individual cell behaviors at the subcellular level with high time resolution, allowing for quantitative dynamic information on cellular processes to be Knowing that the co-expression of YFP-Parkin and KillerRed-dMito is well tolerated by cells, we then tested the use of KillerRed protein for spatiallyencoded ROS generation in living cells using a well-established ROS sensor: 2,7-Dichlorodihydrofluorescein diacetate (DCF).…”