title: Distinct roles of class I and class III phosphatidylinositol 3-kinases in phagosome formation and maturation.
abstract: Phagosomes acquire their microbicidal properties by fusion with lysosomes. Products of phosphatidylinositol 3-kinase (PI 3-kinase) are required for phagosome formation, but their role in maturation is unknown. Using chimeric fluorescent proteins encoding tandem FYVE domains, we found that phosphatidylinositol 3-phosphate (PI[3]P) accumulates greatly but transiently on the phagosomal membrane. Unlike the 3'-phosphoinositides generated by class I PI 3-kinases which are evident in the nascent phagosomal cup, PI(3)P is only detectable after the phagosome has sealed. The class III PI 3-kinase VPS34 was found to be responsible for PI(3)P synthesis and essential for phagolysosome formation. In contrast, selective ablation of class I PI 3-kinase revealed that optimal phagocytosis, but not maturation, requires this type of enzyme. These results highlight the differential functional role of the two families of kinases, and raise the possibility that PI(3)P production by VPS34 may be targeted during the maturation arrest induced by some intracellular parasites.
authors.affiliation: Cell Biology Program, Hospital for Sick Children, Toronto, Ontario M5G 1X8, Canada.
authors.forename: O V
authors.initials: OV
authors.lastname: Vieira
authors.forename: R J
authors.initials: RJ
authors.lastname: Botelho
authors.forename: L
authors.initials: L
authors.lastname: Rameh
authors.forename: S M
authors.initials: SM
authors.lastname: Brachmann
authors.forename: T
authors.initials: T
authors.lastname: Matsuo
authors.forename: H W
authors.initials: HW
authors.lastname: Davidson
authors.forename: A
authors.initials: A
authors.lastname: Schreiber
authors.forename: J M
authors.initials: JM
authors.lastname: Backer
authors.forename: L C
authors.initials: LC
authors.lastname: Cantley
authors.forename: S
authors.initials: S
authors.lastname: Grinstein
mesh_terms: D000730:Androstadienes / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000378:metabolism; D005347:Fibroblasts / Q000378:metabolism; D017930:Genes, Reporter; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008247:Lysosomes / Q000378:metabolism; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D008845:Microinjections; D010587:Phagocytosis / Q000502:physiology*; D010588:Phagosomes / Q000378:metabolism* / Q000648:ultrastructure; D019869:Phosphatidylinositol 3-Kinases / Q000235:genetics / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000077191:Wortmannin
chemical_list: D000730:Androstadienes; D004791:Enzyme Inhibitors; D007074:Immunoglobulin G; D007527:Isoenzymes; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D011993:Recombinant Fusion Proteins; D000077191:Wortmannin