Host cell-dependent secretion and translocation of the LepA and LepB effectors of Legionella pneumophila.
Legionella pneumophila is the Gram-negative bacterial agent of Legionnaires' disease, an acute, often fatal pneumonia. L. pneumophila infects alveolar macrophages, evading the antimicrobial defences of the phagocyte by preventing fusion of the phagosome with lysosomes and avoiding phagosome acidification. The bacteria then modulate the composition of the vacuole so that it takes on the characteristics of the endoplasmic reticulum. Similar events occur when the bacteria infect unicellular protozoa. It is thought that replication in fresh water protozoa provides an environmental reservoir for the organism. Several effector proteins are delivered to the host by the Icm/Dot type IV secretion system (TFSS). Some of these have been shown to participate in the trafficking of the Legionella phagosome. Here we describe the ability of the Icm/Dot TFSS to translocate two effectors, LepA and LepB, that play a role in the non-lytic release of Legionella from protozoa. We report that translocation of the Lep proteins is inhibited by agents that depolymerize actin filaments and that effectors may be secreted into the extracellular medium upon cell contact. Depletion of the Lep proteins by deletion of their genes results in increased ability to lyse red blood cells. In contrast, overexpression of Lep-containing hybrid proteins appears to specifically inhibit the activity of the Icm/Dot TFSS and may prevent the delivery of other effectors that are critical for intracellular multiplication.
Chen
John
J
Department of Microbiology, Columbia University Medical Center, New York, NY 10032, USA.
Reyes
Moraima
M
Clarke
Margaret
M
Shuman
Howard A
HA
D000048:Acanthamoeba / Q000382:microbiology; D000262:Adenylyl Cyclases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D002147:Calmodulin / Q000494:pharmacology; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D004023:Dictyostelium / Q000382:microbiology; D004912:Erythrocytes / Q000382:microbiology; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000382:microbiology; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D020298:U937 Cells
D001426:Bacterial Proteins; D002147:Calmodulin; D002352:Carrier Proteins; C482638:LepA protein, Legionella pneumophila; C482639:LepB protein, Legionella pneumophila; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D000262:Adenylyl Cyclases