title: Synchronizing protein transport in the secretory pathway.
abstract: To be secreted or transported to their target compartments, newly synthesized proteins leave the endoplasmic reticulum to reach the Golgi apparatus, where they are processed and sorted toward their final destinations along the secretory pathway. It is now clear that many Golgi-intersecting and non-intersecting pathways exist in cells to carry out proper transport, modification, and addressing. To analyze and visualize the intracellular trafficking of any secretory protein, we developed the retention using selective hooks (RUSH) system. This assay allows the simultaneous release of a pool of particular secretory proteins from the endoplasmic reticulum and the monitoring of their anterograde trafficking. The use of the RUSH system is detailed in these protocols, from sub-cloning the sequence coding for the protein of interest into RUSH plasmids to visualization of its trafficking.
authors.affiliation: Research Center, Institut Curie, Paris, France.|CNRS UMR144, Paris, France.
authors.forename: Gaelle
authors.initials: G
authors.lastname: Boncompain
authors.affiliation: Research Center, Institut Curie, Paris, France.|CNRS UMR144, Paris, France.
authors.forename: Franck
authors.initials: F
authors.lastname: Perez
mesh_terms: D000818:Animals; D001671:Biochemistry / Q000379:methods*; D002462:Cell Membrane / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D056890:Eukaryota / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D015870:Gene Expression; D017930:Genes, Reporter; D006056:Golgi Apparatus / Q000378:metabolism*; D006801:Humans; D021381:Protein Transport; D055571:Secretory Pathway; D019809:Streptavidin / Q000378:metabolism; D013696:Temperature
chemical_list: D005456:Fluorescent Dyes; D019809:Streptavidin