title: Analysis of protein translocation into the endoplasmic reticulum of human cells.
abstract: The development of small-interfering RNA (siRNA)-mediated gene-silencing strategies has made it possible to study the transport of precursors of soluble and membrane proteins into the endoplasmic reticulum (ER) of human cells. In these approaches, a certain target gene is silenced in the cell type of choice, followed by analysis of the effect of this silencing on the biogenesis of a single or set of precursor polypeptide(s) in cell culture or in cell-free assays involving semi-permeabilized cells and in vitro translations systems. These approaches allow for functional analysis of components of the ER-resident protein transport machinery as well as the elucidation of their potential cell-type variations and regulatory mechanisms. The gene-silencing and subsequent plasmid-based complementation carries the additional benefit of facilitating analysis of the consequences of disease-linked mutations in ER transport components.
authors.affiliation: Medical Biochemistry and Molecular Biology, Saarland University, Homburg, Germany.
authors.forename: Johanna
authors.initials: J
authors.lastname: Dudek
authors.forename: Sven
authors.initials: S
authors.lastname: Lang
authors.forename: Stefan
authors.initials: S
authors.lastname: Schorr
authors.forename: Johannes
authors.initials: J
authors.lastname: Linxweiler
authors.forename: Markus
authors.initials: M
authors.lastname: Greiner
authors.forename: Richard
authors.initials: R
authors.lastname: Zimmermann
mesh_terms: D000818:Animals; D001692:Biological Transport; D018929:Cell Culture Techniques; D002460:Cell Line; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D020868:Gene Silencing; D006801:Humans; D051379:Mice; D010539:Permeability; D021381:Protein Transport; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference
chemical_list: D011506:Proteins