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2015 ; 2015
(ä): 429439
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Activation-Inactivation Cycling of Rab35 and ARF6 Is Required for Phagocytosis of
Zymosan in RAW264 Macrophages
#MMPMID26229970
Egami Y
; Fujii M
; Kawai K
; Ishikawa Y
; Fukuda M
; Araki N
J Immunol Res
2015[]; 2015
(ä): 429439
PMID26229970
show ga
Phagocytosis of zymosan by phagocytes is a widely used model of microbial
recognition by the innate immune system. Live-cell imaging showed that
fluorescent protein-fused Rab35 accumulated in the membranes of phagocytic cups
and then dissociated from the membranes of newly formed phagosomes. By our novel
pull-down assay for Rab35 activity, we found that Rab35 is deactivated
immediately after zymosan internalization into the cells. Phagosome formation was
inhibited in cells expressing the GDP- or GTP-locked Rab35 mutant. Moreover, the
simultaneous expression of ACAP2-a Rab35 effector protein-with GTP-locked Rab35
or the expression of plasma membrane-targeted ACAP2 showed a marked inhibitory
effect on phagocytosis through ARF6 inactivation by the GAP activity of ACAP2.
ARF6, a substrate for ACAP2, was also localized on the phagocytic cups and
dissociated from the membranes of internalized phagosomes. In support of the
microscopic observations, ARF6-GTP pull-down experiments showed that ARF6 is
transiently activated during phagosome formation. Furthermore, the expression of
GDP- or GTP-locked ARF6 mutants also suppresses the uptake of zymosan. These data
suggest that the activation-inactivation cycles of Rab35 and ARF6 are required
for the uptake of zymosan and that ACAP2 is an important component that links
Rab35/ARF6 signaling during phagocytosis of zymosan.