title: Control of protein trafficking by reversible masking of transport signals.
abstract: Systems that allow the control of protein traffic between subcellular compartments have been valuable in elucidating trafficking mechanisms. Most current approaches rely on ligand or light-controlled dimerization, which results in either retardation or enhancement of the transport of a reporter. We developed an alternative approach for trafficking regulation that we term "controlled unmasking of targeting elements" (CUTE). Regulated trafficking is achieved by reversible masking of the signal that directs the reporter to its target organelle, relying on the streptavidin-biotin system. The targeting signal is generated within or immediately after a 38-amino acid streptavidin-binding peptide (SBP) that is appended to the reporter. The binding of coexpressed streptavidin to SBP causes signal masking, whereas addition of biotin causes complex dissociation and triggers protein transport to the target organelle. We demonstrate the application of this approach to the control of nuclear and peroxisomal protein import and the generation of biotin-dependent trafficking through the endocytic and COPI systems. By simultaneous masking of COPI and endocytic signals, we were able to generate a synthetic pathway for efficient transport of a reporter from the plasma membrane to the endoplasmic reticulum.
authors.affiliation: Department of Biology, Technion, Haifa 320003, Israel.
authors.forename: Omer
authors.initials: O
authors.lastname: Abraham
authors.affiliation: Department of Biology, Technion, Haifa 320003, Israel.
authors.forename: Karnit
authors.initials: K
authors.lastname: Gotliv
authors.affiliation: Department of Biology, Technion, Haifa 320003, Israel.
authors.forename: Anna
authors.initials: A
authors.lastname: Parnis
authors.affiliation: Institut Curie, Centre de Recherche, PSL Research University, and CNRS, UMR144, Paris 75248, France.
authors.forename: Gaelle
authors.initials: G
authors.lastname: Boncompain
authors.affiliation: Institut Curie, Centre de Recherche, PSL Research University, and CNRS, UMR144, Paris 75248, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Perez
authors.affiliation: Department of Biology, Technion, Haifa 320003, Israel danc@tx.technion.ac.il.
authors.forename: Dan
authors.initials: D
authors.lastname: Cassel
mesh_terms: D001710:Biotin / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004151:Dipeptides / Q000378:metabolism; D004705:Endocytosis; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D008967:Molecular Biology / Q000379:methods*; D015388:Organelles; D015202:Protein Engineering / Q000379:methods; D021381:Protein Transport / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D019809:Streptavidin / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D004151:Dipeptides; D011993:Recombinant Fusion Proteins; C452700:streptavidin-binding peptide; D001710:Biotin; D019809:Streptavidin; C024575:lysyllysine