The actin-binding protein comitin (p24) is a component of the Golgi apparatus.
Comitin (p24) was first identified in Dictyostelium discoideum as a membrane-associated protein which binds in gel overlay assays to G and F actin. To analyze its actin-binding properties we used purified, bacterially expressed comitin and found that it binds to F actin in spin down experiments and increases the viscosity of F actin solutions even under high-salt conditions. Immunofluorescence studies, cell fractionation experiments and EM studies of vesicles precipitated with comitin-specific monoclonal antibodies showed that comitin was present in D. discoideum on: (a) a perinuclear structure with tubular or fibrillary extensions; and (b) on vesicles distributed throughout the cell. In immunofluorescence experiments using comitin antibodies NIH 3T3 fibroblasts showed a similar staining pattern as D. discoideum cells. Using bona fide Golgi markers the perinuclear structure was identified as the Golgi apparatus. The results were supported by an electron microscopic study using cryosections. Based on these data we propose that also in Dictyostelium the stained perinuclear structure is the Golgi apparatus. In vivo the perinuclear structure was found to be attached to the actin and the microtubule network. Alteration of the actin network or depolymerization of the microtubules led to its dispersal into vesicles distributed throughout the cell. These results suggest that the Golgi apparatus in D. discoideum is connected to the actin network by comitin. This protein seems also to be present in mammalian cells.
Weiner
O H
OH
Max-Planck-Institut für Biochemie, Martinsried, F.R.G.
Murphy
J
J
Griffiths
G
G
Schleicher
M
M
Noegel
A A
AA
D016475:3T3 Cells; D000199:Actins / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D020126:Brefeldin A; D002352:Carrier Proteins; D002451:Cell Compartmentation; D003517:Cyclopentanes / Q000494:pharmacology; D003599:Cytoskeleton / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D004023:Dictyostelium; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D005656:Fungal Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D006056:Golgi Apparatus / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D008840:Microfilament Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism*; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D015739:Nocodazole / Q000494:pharmacology; D015800:Protozoan Proteins; D011994:Recombinant Proteins / Q000378:metabolism
D000199:Actins; D000911:Antibodies, Monoclonal; D000953:Antigens, Protozoan; D002352:Carrier Proteins; D003517:Cyclopentanes; C034037:F-actin-binding proteins; D005656:Fungal Proteins; D008840:Microfilament Proteins; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D020126:Brefeldin A; D015739:Nocodazole; D004121:Dimethyl Sulfoxide