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Published on: July 30, 2014
Actin assembly at membranes controlled by ARF6
D A Schafer1, C D'Souza-Schorey, J A Cooper
1Department of Cell Biology and Physiology, Washington University School of Medicine, St Louis, MO 63110, USA.
Abstract:
The small GTPase, ADP-ribosylation factor-6 (ARF6), has been implicated in regulating membrane traffic and remodeling cortical F-actin. Using real-time video analysis of actin assembly in living cells, we investigated the function and mechanism of ARF6 in control of actin assembly. Expression of an activated form of ARF6 that mimicks the GTP-bound form of the GTPase induced actin assembly resulting in the movement of vesicle-like particles, some of which contain markers for pinosomes. Activated ARF6 also stimulated actin assembly at foci on the ventral surface of the cell and stimulated fluid phase pinocytosis. Particle motility induced by ARF6 involved Arp2/3 complex, tyrosine kinase activity, phospholipase D (PLD) and D3-phosphoinositides, but not phosphatidylinositol 4,5-bisphosphate (PI(4,5)P2). We conclude that ARF6 regulates actin assembly for pinosome motility and at foci on the ventral cell surface.
Insights
ADP-ribosylation factor-6 (ARF6) regulates actin assembly, driving vesicle movement and fluid uptake in cells. This GTPase controls actin remodeling at cell surfaces, impacting cellular processes like pinocytosis.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- ADP-ribosylation factor-6 (ARF6) is a small GTPase involved in membrane traffic and actin cytoskeleton remodeling.
- Understanding ARF6's precise role in actin assembly is crucial for deciphering its cellular functions.
Purpose of the Study:
- To investigate the function and mechanism of ARF6 in controlling actin assembly.
- To elucidate the downstream effectors and signaling pathways regulated by ARF6.
Main Methods:
- Real-time video analysis of actin assembly in living cells.
- Expression of activated ARF6 to mimic its GTP-bound state.
- Analysis of vesicle content and motility.
Main Results:
- Activated ARF6 induced actin assembly, leading to the motility of vesicle-like particles, including pinosomes.
- ARF6 stimulated actin assembly at ventral cell surface foci and enhanced fluid phase pinocytosis.
- ARF6-induced particle motility required Arp2/3 complex, tyrosine kinase activity, phospholipase D (PLD), and D3-phosphoinositides, but not PI(4,5)P2.
Conclusions:
- ARF6 plays a key role in regulating actin assembly for the motility of pinosomes.
- ARF6 controls actin dynamics at specific cellular foci on the ventral cell surface.
- ARF6 integrates signals through PLD and D3-phosphoinositides to modulate actin assembly and membrane trafficking.
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