Role of phosphatidylinositol 3-kinase and Rab5 effectors in phagosomal biogenesis and mycobacterial phagosome maturation arrest

Rutilio A. Fratti, Jonathan M. Backer, Jean Gruenberg, Silvia Corvera, Vojo Deretic

Research output: Contribution to journalArticle

375 Citations (Scopus)

Abstract

Phagosomal biogenesis is a fundamental biological process of particular significance for the function of phagocytic and antigen-presenting cells. The precise mechanisms governing maturation of phagosomes into phagolysosomes are not completely understood. Here, we applied the property of pathogenic mycobacteria to cause phagosome maturation arrest in infected macrophages as a tool to dissect critical steps in phagosomal biogenesis. We report the requirement for 3-phosphoinositides and acquisition of Rab5 effector early endosome autoantigen (EEA1) as essential molecular events necessary for phagosomal maturation. Unlike the model phagosomes containing latex beads, which transiently recruited EEA1, mycobacterial phagosomes excluded this regulator of vesicular trafficking that controls membrane tethering and fusion processes within the endosomal pathway and is recruited to endosomal membranes via binding to phosphatidylinositol 3-phosphate (Ptdlns[3]P). Inhibitors of phosphatidylinositol 3′(OH)-kinase (PI-3K) activity diminished EEA1 recruitment to newly formed latex bead phagosomes and blocked phagosomal acquisition of late endocytic properties, indicating that generation of Ptdlns(3)P plays a role in phagosomal maturation. Microinjection into macrophages of antibodies against EEA1 and the PI-3K hVPS34 reduced acquisition of late endocytic markers by latex bead phagosomes, demonstrating an essential role of these Rab5 effectors in phagosomal biogenesis. The mechanism of EEA1 exclusion from mycobacterial phagosomes was investigated using mycobacterial products. Coating of latex beads with the major mycobacterial cell envelope glycosylated phosphatidylinositol lipoarabinomannan isolated from the virulent Mycobacterium tuberculosis H37Rv, inhibited recruitment of EEA1 to latex bead phagosomes, and diminished their maturation. These findings define the generation of phosphatidylinositol 3-phosphate and EEA1 recruitment as: (a) important regulatory events in phagosomal maturation and (b) critical molecular targets affected by M. tuberculosis. This study also identifies mycobacterial phosphoinositides as products with specialized toxic properties, interfering with discrete trafficking stages in phagosomal maturation.

Original languageEnglish (US)
Pages (from-to)631-644
Number of pages14
JournalJournal of Cell Biology
Volume154
Issue number3
DOIs
StatePublished - Aug 6 2001

Fingerprint

Phosphatidylinositol 3-Kinase
Phagosomes
Microspheres
Phosphatidylinositol 3-Kinases
Phosphatidylinositols
Mycobacterium tuberculosis
Macrophages
Biological Phenomena
Glycosylphosphatidylinositols
Membrane Fusion
Poisons
Endosomes
Autoantigens
Microinjections
Antigen-Presenting Cells
Mycobacterium

Keywords

  • EEA1
  • Endosome
  • hVPS34
  • LAM
  • LBPA

ASJC Scopus subject areas

  • Cell Biology

Cite this

Role of phosphatidylinositol 3-kinase and Rab5 effectors in phagosomal biogenesis and mycobacterial phagosome maturation arrest. / Fratti, Rutilio A.; Backer, Jonathan M.; Gruenberg, Jean; Corvera, Silvia; Deretic, Vojo.

In: Journal of Cell Biology, Vol. 154, No. 3, 06.08.2001, p. 631-644.

Research output: Contribution to journalArticle

Fratti, Rutilio A. ; Backer, Jonathan M. ; Gruenberg, Jean ; Corvera, Silvia ; Deretic, Vojo. / Role of phosphatidylinositol 3-kinase and Rab5 effectors in phagosomal biogenesis and mycobacterial phagosome maturation arrest. In: Journal of Cell Biology. 2001 ; Vol. 154, No. 3. pp. 631-644.
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