title: The Legionella pneumophila effector VipA is an actin nucleator that alters host cell organelle trafficking.
abstract: Legionella pneumophila, the causative agent of Legionnaires' disease, invades and replicates within macrophages and protozoan cells inside a vacuole. The type IVB Icm/Dot secretion system is necessary for the translocation of effector proteins that modulate vesicle trafficking pathways in the host cell, thus avoiding phagosome-lysosome fusion. The Legionella VipA effector was previously identified by its ability to interfere with organelle trafficking in the Multivesicular Body (MVB) pathway when ectopically expressed in yeast. In this study, we show that VipA binds actin in vitro and directly polymerizes microfilaments without the requirement of additional proteins, displaying properties distinct from other bacterial actin nucleators. Microscopy studies revealed that fluorescently tagged VipA variants localize to puncta in eukaryotic cells. In yeast these puncta are associated with actin-rich regions and components of the Multivesicular Body pathway such as endosomes and the MVB-associated protein Bro1. During macrophage infection, native translocated VipA associated with actin patches and early endosomes. When ectopically expressed in mammalian cells, VipA-GFP displayed a similar distribution ruling out the requirement of additional effectors for binding to its eukaryotic targets. Interestingly, a mutant form of VipA, VipA-1, that does not interfere with organelle trafficking is also defective in actin binding as well as association with early endosomes and shows a homogeneous cytosolic localization. These results show that the ability of VipA to bind actin is related to its association with a specific subcellular location as well as its role in modulating organelle trafficking pathways. VipA constitutes a novel type of actin nucleator that may contribute to the intracellular lifestyle of Legionella by altering cytoskeleton dynamics to target host cell pathways.
authors.affiliation: Department of Microbiology and Immunology, Columbia University Medical Center, New York, New York, USA. irinafranco@hotmail.com
authors.forename: Irina Saraiva
authors.initials: IS
authors.lastname: Franco
authors.forename: Nadim
authors.initials: N
authors.lastname: Shohdy
authors.forename: Howard A
authors.initials: HA
authors.lastname: Shuman
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000502:physiology*; D001692:Biological Transport / Q000235:genetics / Q000502:physiology; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D054884:Host-Pathogen Interactions / Q000235:genetics / Q000502:physiology; D006801:Humans; D016952:Legionella pneumophila / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity / Q000502:physiology; D016296:Mutagenesis / Q000502:physiology; D015388:Organelles / Q000378:metabolism* / Q000502:physiology; D030781:Organisms, Genetically Modified; D012441:Saccharomyces cerevisiae; D020298:U937 Cells
chemical_list: D001426:Bacterial Proteins