PMID: 9450963Apr 4, 1998Paper

Involvement of ATP-dependent Pseudomonas exotoxin translocation from a late recycling compartment in lymphocyte intoxication procedure

Molecular Biology of the Cell
M AlamiB Beaumelle

Abstract

Pseudomonas exotoxin (PE) is a cytotoxin which, after endocytosis, is delivered to the cytosol where it inactivates protein synthesis. Using diaminobenzidine cytochemistry, we found over 94% of internalized PE in transferrin (Tf) -positive endosomes of lymphocytes. When PE translocation was examined in a cell-free assay using purified endocytic vesicles, more than 40% of endosomal 125I-labeled PE was transported after 2 h at 37 degrees C, whereas a toxin inactivated by point mutation in its translocation domain was not translocated. Sorting of endosomes did not allow cell-free PE translocation, whereas active PE transmembrane transport was observed after > 10 min of endocytosis when PE and fluorescent-Tf were localized by confocal immunofluorescence microscopy within a rab5-positive and rab4- and rab7-negative recycling compartment in the pericentriolar region of the cell. Accordingly, when PE delivery to this structure was inhibited using a 20 degrees C endocytosis temperature, subsequent translocation from purified endosomes was impaired. Translocation was also inhibited when endosomes were obtained from cells labeled with PE in the presence of brefeldin A, which caused fusion of translocation-competent recycling endosomes wi...Continue Reading

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Citations

Mar 21, 2008·Journal of Biomedical Science·Diana V PastranaDavid J Fitzgerald
Apr 22, 2009·Infection and Immunity·Juliette Morlon-GuyotBruno Beaumelle
Mar 8, 2013·PLoS Pathogens·Daniela S SilvaNuno M S dos Santos
Dec 10, 2009·Cell Biology International·Fabienne RayneBruno Beaumelle
Apr 11, 2001·Molecular Biology of the Cell·J S BergR E Cheney
Mar 19, 2020·Cellular Microbiology·Francisco Sarmento MesquitaOksana A Sergeeva
Mar 4, 2003·The Journal of Biological Chemistry·Juliette Morlon-GuyotBruno Beaumelle
Apr 23, 2015·Bioorganicheskaia khimiia·A N Noskov
Mar 22, 2007·Journal of Cell Science·Montserrat RodríguezBruno Beaumelle

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