title: Characterization of Salmonella-induced filaments (Sifs) reveals a delayed interaction between Salmonella-containing vacuoles and late endocytic compartments.
abstract: Salmonella typhimurium is a facultative intracellular pathogen that colonizes host cells throughout the course of infection. A unique feature of this pathogen is its ability to enter into (invade) epithelial cells and elongate the vacuole within which it resides into tubular structures called Salmonella-induced filaments (Sifs). In this study we sought to characterize the mechanism of Sif formation by immunofluorescence analysis using subcellular markers. The late endosomal lipid lysobisphosphatidic acid associated in a punctate pattern with the Salmonella-containing vacuole, starting 90 min after infection and increasing thereafter. Lysobisphosphatidic acid-rich vesicles were also found to interact with Sifs, at numerous sites along the tubules. Similarly, cholesterol-rich vesicles were also found in association with intracellular bacteria and Sifs. The lysosomal hydrolase cathepsin D was present in Sifs, both in a punctate pattern and, at later times, predominantly in an uninterrupted linear pattern. Rab7 associated with Sifs and expression of the N125I dominant negative mutant of this GTPase inhibited Sif formation. Transfection of HeLa cells with a vector encoding SifA fused to the green fluorescent protein caused swelling and aggregation of lysobisphosphatidic acid-containing compartments, suggesting that this virulence factor directs membrane fusion events involving late endosomes. Our findings demonstrate that Sif formation involves fusion of late endocytic compartments with the Salmonella-containing vacuole, and suggest that SifA modulates this event.
authors.affiliation: Biotechnology Laboratory and Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, V6T-1Z3, Canada.
authors.forename: J H
authors.initials: JH
authors.lastname: Brumell
authors.forename: P
authors.initials: P
authors.lastname: Tang
authors.forename: S D
authors.initials: SD
authors.lastname: Mills
authors.forename: B B
authors.initials: BB
authors.lastname: Finlay
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001426:Bacterial Proteins; D001692:Biological Transport; D002402:Cathepsin D / Q000378:metabolism / Q000494:pharmacology; D002784:Cholesterol / Q000378:metabolism; D004705:Endocytosis; D011992:Endosomes / Q000378:metabolism; D020558:GTP Phosphohydrolases / Q000378:metabolism; D005799:Genes, Dominant; D006023:Glycoproteins / Q000378:metabolism*; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D051907:Lysosomal Membrane Proteins; D008264:Macrophages / Q000378:metabolism / Q000382:microbiology; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009154:Mutation; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012486:Salmonella typhimurium / Q000472:pathogenicity*; D013997:Time Factors; D014162:Transfection; D020691:rab GTP-Binding Proteins / Q000378:metabolism; D000090802:rab7 GTP-Binding Proteins
chemical_list: D015703:Antigens, CD; D001426:Bacterial Proteins; D006023:Glycoproteins; D008164:Luminescent Proteins; D051907:Lysosomal Membrane Proteins; D008562:Membrane Glycoproteins; C421862:SifA protein, Salmonella; D000090802:rab7 GTP-Binding Proteins; C000716463:rab7 GTP-binding proteins, human; C000716465:rab7 GTP-binding proteins, mouse; D049452:Green Fluorescent Proteins; D002784:Cholesterol; D002402:Cathepsin D; D020558:GTP Phosphohydrolases; D020691:rab GTP-Binding Proteins