Regulation of FcgammaR-stimulated phagocytosis by the 72-kDa inositol polyphosphate 5-phosphatase: SHIP1, but not the 72-kDa 5-phosphatase, regulates complement receptor 3 mediated phagocytosis by differential recruitment of these 5-phosphatases to the phagocytic cup.
Macrophages phagocytose particles to resolve infections and remove apoptotic cells. Phosphoinositide 3-kinase generates phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P(3)] is restricted to the phagocytic cup, promoting phagocytosis. The PtdIns(3,4,5)P(3) 5-phosphatase (5-ptase) Src homology 2 (SH2) domain-containing inositol-5-phosphatase 1 (SHIP1) inhibits phagocytosis. We report here that another PtdIns(3,4,5)P(3)-5-ptase, the 72-kDa-5-phosphatase (72-5ptase), inhibits Fcgamma receptor (FcgammaR)- but not complement receptor 3 (CR3)-mediated phagocytosis, affecting pseudopod extension and phagosome closure. In contrast, SHIP1 inhibited FcgammaR and CR3 phagocytosis with greater effects on CR3-stimulated phagocytosis. The 72-5ptase and SHIP1 were both dynamically recruited to FcgammaR-stimulated phagocytic cups, but only SHIP1 was recruited to CR3-stimulated phagocytic cups. To determine whether 5-ptases focally degrade PtdIns(3,4,5)P(3) at the phagocytic cup after specific stimuli, time-lapse imaging of specific biosensors was performed. Transfection of dominant-negative 72-5ptase or 72-5ptase small interfering RNA (siRNA) resulted in amplified and prolonged PtdIns(3,4,5)P(3) at the phagocytic cup in response to FcgammaR- but not CR3-stimulation. In contrast, macrophages from Ship1(-/-)/AktPH-GFP transgenic mice exhibited increased and sustained PtdIns(3,4,5)P(3) at the cup in response to CR3 activation, with minimal changes to FcgammaR activation. Therefore, 72-5ptase and SHIP1 exhibit specificity in regulating FcgammaR- versus CR3-stimulated phagocytosis by controlling the amplitude and duration of PtdIns(3,4,5)P(3) at the phagocytic cup.
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Australia.
Kristy A
KA
Horan
Ken-Ichi
K
Watanabe
Anne M
AM
Kong
Charles G
CG
Bailey
John E J
JE
Rasko
Takehiko
T
Sasaki
Christina A
CA
Mitchell
D000818:Animals; D002460:Cell Line; D000072181:Inositol Polyphosphate 5-Phosphatases; D016177:Macrophage-1 Antigen / Q000502:physiology*; D008264:Macrophages / Q000201:enzymology / Q000276:immunology* / Q000648:ultrastructure; D051379:Mice; D008822:Mice, Transgenic; D010587:Phagocytosis / Q000276:immunology*; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases; D010744:Phosphoric Monoester Hydrolases / Q000172:deficiency / Q000502:physiology*; D021381:Protein Transport; D011554:Pseudopodia; D017452:Receptors, IgG / Q000276:immunology*; D000099182:Phosphoinositide 5-Phosphatases
D000072181:Inositol Polyphosphate 5-Phosphatases; D016177:Macrophage-1 Antigen; D000072183:Phosphatidylinositol-3,4,5-Trisphosphate 5-Phosphatases; D010744:Phosphoric Monoester Hydrolases; D017452:Receptors, IgG; D000099182:Phosphoinositide 5-Phosphatases; C000606432:Inpp5a protein, mouse; C000606433:Inpp5d protein, mouse