title: A light-triggered protein secretion system.
abstract: Optical control of protein interactions has emerged as a powerful experimental paradigm for manipulating and studying various cellular processes. Tools are now available for controlling a number of cellular functions, but some fundamental processes, such as protein secretion, have been difficult to engineer using current optical tools. Here we use UVR8, a plant photoreceptor protein that forms photolabile homodimers, to engineer the first light-triggered protein secretion system. UVR8 fusion proteins were conditionally sequestered in the endoplasmic reticulum, and a brief pulse of light triggered robust forward trafficking through the secretory pathway to the plasma membrane. UVR8 was not responsive to excitation light used to image cyan, green, or red fluorescent protein variants, allowing multicolor visualization of cellular markers and secreted protein cargo as it traverses the cellular secretory pathway. We implemented this novel tool in neurons to demonstrate restricted, local trafficking of secretory cargo near dendritic branch points.
authors.affiliation: Department of Pharmacology, University of Colorado Denver School of Medicine, Aurora, CO 80045, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Chen
authors.forename: Emily S
authors.initials: ES
authors.lastname: Gibson
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Kennedy
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D029681:Arabidopsis Proteins / Q000378:metabolism; D019556:COS Cells; D002462:Cell Membrane / Q000378:metabolism; D002522:Chlorocebus aethiops; D002868:Chromosomal Proteins, Non-Histone / Q000378:metabolism; D003712:Dendrites / Q000378:metabolism; D017343:Genes, Plant; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D008027:Light; D008856:Microscopy, Fluorescence; D009474:Neurons / Q000378:metabolism; D055095:Optics and Photonics / Q000379:methods*; D025941:Protein Interaction Mapping / Q000379:methods; D021381:Protein Transport / Q000528:radiation effects; D011506:Proteins / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D014466:Ultraviolet Rays
chemical_list: D029681:Arabidopsis Proteins; D002868:Chromosomal Proteins, Non-Histone; D011506:Proteins; C525052:Uvr8 protein, Arabidopsis; D049452:Green Fluorescent Proteins