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Analysis of Slow-cycling Variants of the Light-inducible Nuclear Protein Export System LEXY in Mammalian Cells
Giada Forlani
Enoch Antwi
Daniel Georg Weis
Mehmet Ali Öztürk
Bastian A.W Queck
Dominik Brecht
Barbara Di Ventura
出版
Universität
, 2022
URL
http://books.google.com.hk/books?id=-YaWzwEACAAJ&hl=&source=gbs_api
註釋
Abstract: The optogenetic tool LEXY consists of the second light oxygen voltage (LOV) domain of Avena sativa phototropin 1 mutated to contain a nuclear export signal. It allows exporting from the nucleus with blue light proteins of interest (POIs) genetically fused to it. Mutations slowing the dark recovery rate of the LOV domain within LEXY were recently shown to allow for better depletion of some POIs from the nucleus in Drosophila embryos and for the usage of low light illumination regimes. We investigated these variants in mammalian cells and found they increase the cytoplasmic localization of the proteins we tested after illumination, but also during the dark phases, which corresponds to higher leakiness of the system. These data suggest that, when aiming to sequester into the nucleus a protein with a cytoplasmic function, the original LEXY is preferable. The iLEXY variants are, instead, advantageous when wanting to deplete the nucleus of the POI as much as possible