title: Use of RNA interference in Drosophila S2 cells to identify host pathways controlling compartmentalization of an intracellular pathogen.
abstract: Three genome-wide RNA interference screens were performed in Drosophila S2 cells to dissect the contribution of host processes to Listeria monocytogenes entry, vacuolar escape, and intracellular growth. Among the 116 genes identified, several host pathways previously unrecognized as playing a role in listerial pathogenesis were identified: knockdowns affecting vacuolar trafficking to and from the multivesicular body bypassed the requirement for the essential pore-forming toxin listeriolysin O in mediating escape from phagocytic vacuoles and knockdowns affecting either subunit of serine palmitoyltransferase, a key enzyme in ceramide and sphingolipid biosynthesis, enhanced the toxicity of listeriolysin O expressed in the host cell cytosol, leading to lack of appropriate toxin activity compartmentalization and host cell death. Genome-wide RNA interference screens using Drosophila S2 cells proved to be a powerful approach to dissect host-pathogen interactions.
authors.affiliation: Department of Molecular and Cell Biology and the School of Public Health, 508 Barker Hall, University of California, Berkeley, CA 94720-3202, USA.
authors.forename: Luisa W
authors.initials: LW
authors.lastname: Cheng
authors.forename: Julie P M
authors.initials: JP
authors.lastname: Viala
authors.forename: Nico
authors.initials: N
authors.lastname: Stuurman
authors.forename: Ursula
authors.initials: U
authors.lastname: Wiedemann
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Vale
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Portnoy
mesh_terms: D000818:Animals; D001427:Bacterial Toxins / Q000378:metabolism; D002460:Cell Line; D003593:Cytoplasm / Q000378:metabolism* / Q000382:microbiology; D004330:Drosophila; D020869:Gene Expression Profiling; D006360:Heat-Shock Proteins / Q000378:metabolism; D006460:Hemolysin Proteins; D008089:Listeria monocytogenes / Q000378:metabolism; D008088:Listeriosis / Q000235:genetics / Q000378:metabolism*; D010588:Phagosomes / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D034622:RNA Interference
chemical_list: D001427:Bacterial Toxins; D006360:Heat-Shock Proteins; D006460:Hemolysin Proteins; C038705:hlyA protein, Listeria monocytogenes