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Rab35 regulates phagosome formation throughrecruitment of ACAP2 in macrophages duringFccR-mediated phagocytosis

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Rab35 regulates phagosome formation throughrecruitment of ACAP2 in macrophages duringFccR-mediated phagocytosis

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Phagosome formation and subsequent maturation are complex sequences of events that involve actin cytoskeleton remodeling and
membrane trafficking. Here, we demonstrate that the Ras-related protein Rab35 is involved in the early stage of FccR-mediated
phagocytosis in macrophages. Live-cell image analysis revealed that Rab35 was markedly concentrated at the membrane where IgGopsonized
erythrocytes (IgG-Es) are bound. Rab35 silencing by RNA interference (RNAi) or the expression of GDP- or GTP-locked
Rab35 mutant drastically reduced the rate of phagocytosis of IgG-Es. Actin-mediated pseudopod extension to form phagocytic cups
was disturbed by the Rab35 silencing or the expression of GDP-Rab35, although initial actin assembly at the IgG-E binding sites was
not inhibited. Furthermore, GTP-Rab35-dependent recruitment of ACAP2, an ARF6 GTPase-activating protein, was shown in the
phagocytic cup formation. Concomitantly, overexpression of ACAP2 along with GTP-locked Rab35 showed a synergistic inhibitory
effect on phagocytosis. It is likely that Rab35 regulates actin-

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Rab35 regulates Arf6 activity through centaurin-b2(ACAP2) during neurite outgrowth  (essential factors for neurite outgrowth)

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Rab35 regulates Arf6 activity through centaurin-b2(ACAP2) during neurite outgrowth  (essential factors for neurite outgrowth)

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centaurin-b2 (also known as ACAP2) functions both as a Rab35 effector and as an Arf6-GTPase-activating protein (GAP) during neurite outgrowth of PC12 cells.
Rab35 accumulates at Arf6-positive endosomes in response to nerve growth factor (NGF) stimulation and that centaurin-b2 is recruited to the same compartment in a Rab35-dependent manner. We further showed by knockdown and rescue experiments that after the Rab35-dependent recruitment of centaurin-b2, the Arf6-GAP activity of centaurin-b2 at the Arf6-positive endosomes was indispensable for NGF-induced neurite outgrowth. These findings suggest a novel mode of crosstalk between Rab and Arf: a Rab effector and Arf-GAP coupling mechanism, in which Arf-GAP is recruited to a specific membrane compartment by its Rab effector
function

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Human CNT-1 Homolog ACAP2has a Pro-Apoptotic Role inCancer Cells

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Human CNT-1 Homolog ACAP2has a Pro-Apoptotic Role inCancer Cells

# data[].attributes.body
ACAP2 is a demonstratedRAB35 effector and a GTPase activatingprotein (GAP) for ARF6.24-26 Wefocused our studies on ACAP2 andACAP3, as these proteins display themost significant homology with CNT-1(33% identical, 50% similar and 35%identical, 53% similar, respectively), especiallywithin their PH domains (52%identical, 75% similar and 52% identical,76% similar, respectively, Fig. 1B),which is the critical domain for tCNT-1-mediated AKT inactivation.

# data[].attributes.body
sarahm.jun@mail.utoronto.ca

lily.trinh@mail.utoronto.ca

adnan.sharif@mail.utoronto.ca

tingixn.xiao@mail.utoronto.ca

Topic: ACAP2 gene regulations and implications in ZGA and MZT