A Fluorescent Live Imaging Screening Assay Based on Translocation Criteria Identifies Novel Cytoplasmic Proteins Implicated in G Protein-coupled Receptor Signaling Pathways.

Molecular & Cellular Proteomics : MCP
Sandra LecatJean-Luc Galzi

Abstract

Several cytoplasmic proteins that are involved in G protein-coupled receptor signaling cascades are known to translocate to the plasma membrane upon receptor activation, such as beta-arrestin2. Based on this example and in order to identify new cytoplasmic proteins implicated in the ON-and-OFF cycle of G protein-coupled receptor, a live-imaging screen of fluorescently labeled cytoplasmic proteins was performed using translocation criteria. The screening of 193 fluorescently tagged human proteins identified eight proteins that responded to activation of the tachykinin NK2 receptor by a change in their intracellular localization. Previously we have presented the functional characterization of one of these proteins, REDD1, that translocates to the plasma membrane. Here we report the results of the entire screening. The process of cell activation was recorded on videos at different time points and all the videos can be visualized on a dedicated website. The proteins BAIAP3 and BIN1, partially translocated to the plasma membrane upon activation of NK2 receptors. Proteins ARHGAP12 and PKM2 translocated toward membrane blebs. Three proteins that associate with the cytoskeleton were of particular interest : PLEKHH2 rearranged from indi...Continue Reading

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Citations

Nov 18, 2016·Journal of Cell Science·Elisabetta Argenzio, Wouter H Moolenaar
Dec 26, 2018·Circulation Research·Vahitha B Abdul-SalamBeata Wojciak-Stothard
Mar 31, 2018·The Biochemical Journal·Gualtiero AlvisiAnnalisa Radeghieri
Jun 19, 2019·Nature Cell Biology·Jessica DiringRichard Treisman
Sep 3, 2020·American Journal of Physiology. Cell Physiology·Florian A BrittoFrancois B Favier
Nov 6, 2020·Brain, Behavior, and Immunity·Seyma KatrinliAlicia K Smith

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