Activated ADP-ribosylation factor assembles distinct pools of actin on golgi membranes.
The small GTP-binding protein ADP-ribosylation factor (ARF) has been shown to regulate the interaction of actin and actin-binding proteins with the Golgi apparatus. Here we report that ARF activation stimulates the assembly of distinct pools of actin on Golgi membranes. One pool of actin cofractionates with coatomer (COPI)- coated vesicles and is sensitive to salt extraction and the plus end actin-binding toxin cytochalasin D. A second ARF-dependent actin pool remains on the Golgi membranes following vesicle extraction and is insensitive to cytochalasin D. Isolation of the salt-extractable ARF-dependent actin from the Golgi reveals that it is bound to a distinct repertoire of actin-binding proteins. The two abundant actin-binding proteins of the ARF-dependent actin complex are identified as spectrin and drebrin. We show that drebrin is a specific component of the cytochalasin D-sensitive, ARF-dependent actin pool on the Golgi. Finally, we show that depolymerization of this actin pool with cytochalasin D increases the extent of the salt-dependent release of COPI-coated vesicles from the Golgi following cell-free budding reactions. Together these data suggest that regulation of the actin-based cytoskeleton may play an important role during ARF-mediated transport vesicle assembly or release on the Golgi.
Fucini
R V
RV
Department of Physiology and Biophysics, University of Iowa College of Medicine, Iowa City, Iowa 52242, USA.
Navarrete
A
A
Vadakkan
C
C
Lacomis
L
L
Erdjument-Bromage
H
H
Tempst
P
P
Stamnes
M
M
D020727:ADP-Ribosylation Factors / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D000818:Animals; D006056:Golgi Apparatus / Q000378:metabolism*; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D007425:Intracellular Membranes / Q000378:metabolism*; D009479:Neuropeptides / Q000378:metabolism; D051381:Rats
D000199:Actins; D009479:Neuropeptides; C053386:drebrins; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D020727:ADP-Ribosylation Factors