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.jpg): Failed to open stream: No such file or directory in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 117 J+Immunol
2017 ; 199
(1
): 271-277
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Golgi-Associated Protein Kinase C-? Is Delivered to Phagocytic Cups: Role of
Phosphatidylinositol 4-Phosphate
#MMPMID28539432
Hanes CM
; D'Amico AE
; Ueyama T
; Wong AC
; Zhang X
; Hynes WF
; Barroso MM
; Cady NC
; Trebak M
; Saito N
; Lennartz MR
J Immunol
2017[Jul]; 199
(1
): 271-277
PMID28539432
show ga
Protein kinase C-? (PKC-?) at phagocytic cups mediates the membrane fusion
necessary for efficient IgG-mediated phagocytosis. The C1B and pseudosubstrate
(?PS) domains are necessary and sufficient for this concentration. C1B binds
diacylglycerol; the docking partner for ?PS is unknown. Liposome assays revealed
that the ?PS binds phosphatidylinositol 4-phosphate (PI4P) and PI(3,5)P(2)
Wortmannin, but not LY294002, inhibits PKC-? concentration at cups and
significantly reduces the rate of phagocytosis. As Wortmannin inhibits PI4
kinase, we hypothesized that PI4P mediates the PKC-? concentration at cups and
the rate of phagocytosis. PKC-? colocalizes with the trans-Golgi network (TGN)
PI4P reporter, P4M, suggesting it is tethered at the TGN. Real-time imaging of
GFP-PKC-?-expressing macrophages revealed a loss of Golgi-associated PKC-? during
phagocytosis, consistent with a Golgi-to-phagosome translocation. Treatment with
PIK93, a PI4 kinase inhibitor, reduces PKC-? at both the TGN and the cup,
decreases phagocytosis, and prevents the increase in capacitance that accompanies
membrane fusion. Finally, expression of the Golgi-directed PI4P phosphatase,
hSac1-K2A, recapitulates the PIK93 phenotype, confirming that Golgi-associated
PI4P is critical for efficient phagocytosis. Together these data are consistent
with a model in which PKC-? is tethered to the TGN via an ?PS-PI4P interaction.
The TGN-associated pool of PKC-? concentrates at the phagocytic cup where it
mediates the membrane fusion necessary for phagocytosis. The novelty of these
data lies in the demonstration that ?PS binds PI4P and PI(3,5)P(2) and that PI4P
is necessary for PKC-? localization at the TGN, its translocation to the
phagocytic cup, and the membrane fusion required for efficient Fc [?]
receptor-mediated phagocytosis.
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