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.
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1 October 2001
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October 01 2001
Distinct roles of class I and class III phosphatidylinositol 3-kinases in phagosome formation and maturation
Otilia V. Vieira,
Otilia V. Vieira
1Cell Biology Program, Hospital for Sick Children and Department of Biochemistry, University of Toronto, Ontario M5G 1X8, Canada
2Center for Neurosciences, University of Coimbra, 3000 Coimbra, Portugal
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Roberto J. Botelho,
Roberto J. Botelho
1Cell Biology Program, Hospital for Sick Children and Department of Biochemistry, University of Toronto, Ontario M5G 1X8, Canada
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Lucia Rameh,
Lucia Rameh
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
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Saskia M. Brachmann,
Saskia M. Brachmann
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
4Freie Universitaet Berlin, Institut fuer Biochemie, 14195 Berlin, Germany
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Tsuyoshi Matsuo,
Tsuyoshi Matsuo
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
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Howard W. Davidson,
Howard W. Davidson
5Wellcome Trust Center for Molecular Mechanisms of Disease, University of Cambridge, Addenbrookes Hospital, Cambridge CB2 2QQ, UK
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Alan Schreiber,
Alan Schreiber
6Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104
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Jonathan M. Backer,
Jonathan M. Backer
7Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10032
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Lewis C. Cantley,
Lewis C. Cantley
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
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Sergio Grinstein
Sergio Grinstein
1Cell Biology Program, Hospital for Sick Children and Department of Biochemistry, University of Toronto, Ontario M5G 1X8, Canada
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Otilia V. Vieira
1Cell Biology Program, Hospital for Sick Children and Department of Biochemistry, University of Toronto, Ontario M5G 1X8, Canada
2Center for Neurosciences, University of Coimbra, 3000 Coimbra, Portugal
Roberto J. Botelho
1Cell Biology Program, Hospital for Sick Children and Department of Biochemistry, University of Toronto, Ontario M5G 1X8, Canada
Lucia Rameh
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
Saskia M. Brachmann
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
4Freie Universitaet Berlin, Institut fuer Biochemie, 14195 Berlin, Germany
Tsuyoshi Matsuo
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
Howard W. Davidson
5Wellcome Trust Center for Molecular Mechanisms of Disease, University of Cambridge, Addenbrookes Hospital, Cambridge CB2 2QQ, UK
Alan Schreiber
6Department of Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104
Jonathan M. Backer
7Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, NY 10032
Lewis C. Cantley
3Divison of Signal Transduction, Beth Israel Deaconess Medical Center, Harvard Institutes of Medicine, Boston, MA 02115
Sergio Grinstein
1Cell Biology Program, Hospital for Sick Children and Department of Biochemistry, University of Toronto, Ontario M5G 1X8, Canada
Address correspondence to Sergio Grinstein, Division of Cell Biology, Hospital for Sick Children, 555 University Ave., Toronto, Ontario M5G 1X8, Canada. Tel.: (416) 813-5727. Fax: (416) 813-5028. E-mail: [email protected]
*
Abbreviations used in this paper: DIC, differential interference contrast; GFP, green fluorescent protein; PI 3-kinase, phosphatidylinositol 3-kinase; PI(3)P, phosphatidylinositol 3-phosphate; RBC, red blood cells.
Received:
July 18 2001
Revision Received:
August 29 2001
Accepted:
August 29 2001
Online ISSN: 1540-8140
Print ISSN: 0021-9525
The Rockefeller University Press
2001
J Cell Biol (2001) 155 (1): 19–26.
Article history
Received:
July 18 2001
Revision Received:
August 29 2001
Accepted:
August 29 2001
Citation
Otilia V. Vieira, Roberto J. Botelho, Lucia Rameh, Saskia M. Brachmann, Tsuyoshi Matsuo, Howard W. Davidson, Alan Schreiber, Jonathan M. Backer, Lewis C. Cantley, Sergio Grinstein; Distinct roles of class I and class III phosphatidylinositol 3-kinases in phagosome formation and maturation . J Cell Biol 1 October 2001; 155 (1): 19–26. doi: https://doi.org/10.1083/jcb.200107069
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