title: Subcellular localization of legionella Dot/Icm effectors.
abstract: The translocation of effector proteins by the Dot/Icm type IV secretion system is central to the ability of Legionella pneumophila to persist and replicate within eukaryotic cells. The subcellular localization of translocated Dot/Icm proteins in host cells provides insight into their function. Through co-staining with host cell markers, effector proteins may be localized to specific subcellular compartments and membranes, which frequently reflects their host cell target and mechanism of action. In this chapter, we describe protocols to (1) localize effector proteins within cells by ectopic expression using green fluorescent protein fusions and (2) localize effector proteins within infected cells using epitope-tagged effector proteins and immuno-fluorescence microscopy.
authors.affiliation: Department of Microbiology and Immunology, University of Melbourne, Melbourne, VIC, Australia.
authors.forename: Adam J
authors.initials: AJ
authors.lastname: Vogrin
authors.forename: Aurelie
authors.initials: A
authors.lastname: Mousnier
authors.forename: Gad
authors.initials: G
authors.lastname: Frankel
authors.forename: Elizabeth L
authors.initials: EL
authors.lastname: Hartland
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D058947:Bacterial Secretion Systems; D003001:Cloning, Molecular; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007875:Legionella / Q000235:genetics / Q000378:metabolism*; D016952:Legionella pneumophila / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
chemical_list: D001426:Bacterial Proteins; D058947:Bacterial Secretion Systems; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins