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J Cell Biol. 1993 October 1; 123(1): 23–34.
PMCID: PMC2119810

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.

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Selected References

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  • Adams RJ, Pollard TD. Membrane-bound myosin-I provides new mechanisms in cell motility. Cell Motil Cytoskeleton. 1989;14(2):178–182. [PubMed]
  • Allen RD, Metuzals J, Tasaki I, Brady ST, Gilbert SP. Fast axonal transport in squid giant axon. Science. 1982 Dec 10;218(4577):1127–1129. [PubMed]
  • Allen RD, Weiss DG, Hayden JH, Brown DT, Fujiwake H, Simpson M. Gliding movement of and bidirectional transport along single native microtubules from squid axoplasm: evidence for an active role of microtubules in cytoplasmic transport. J Cell Biol. 1985 May;100(5):1736–1752. [PMC free article] [PubMed]
  • Bauduin H, Stock C, Vincent D, Grenier JF. Microfilamentous system and secretion of enzyme in the exocrine pancreas. Effect of cytochalasin B. J Cell Biol. 1975 Jul;66(1):165–181. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brady ST, Lasek RJ, Allen RD, Yin HL, Stossel TP. Gelsolin inhibition of fast axonal transport indicates a requirement for actin microfilaments. Nature. 1984 Jul 5;310(5972):56–58. [PubMed]
  • Cardelli JA, Golumbeski GS, Woychik NA, Ebert DL, Mierendorf RC, Dimond RL. Defining the intracellular localization pathways followed by lysosomal enzymes in Dictyostelium discoideum. Methods Cell Biol. 1987;28:139–155. [PubMed]
  • Claviez M, Pagh K, Maruta H, Baltes W, Fisher P, Gerisch G. Electron microscopic mapping of monoclonal antibodies on the tail region of Dictyostelium myosin. EMBO J. 1982;1(8):1017–1022. [PubMed]
  • Coudrier E, Durrbach A, Louvard D. Do unconventional myosins exert functions in dynamics of membrane compartments? FEBS Lett. 1992 Jul 27;307(1):87–92. [PubMed]
  • Döring V, Schleicher M, Noegel AA. Dictyostelium annexin VII (synexin). cDNA sequence and isolation of a gene disruption mutant. J Biol Chem. 1991 Sep 15;266(26):17509–17515. [PubMed]
  • Duden R, Griffiths G, Frank R, Argos P, Kreis TE. Beta-COP, a 110 kd protein associated with non-clathrin-coated vesicles and the Golgi complex, shows homology to beta-adaptin. Cell. 1991 Feb 8;64(3):649–665. [PubMed]
  • Goldberg DJ, Harris DA, Lubit BW, Schwartz JH. Analysis of the mechanism of fast axonal transport by intracellular injection of potentially inhibitory macromolecules: evidence for a possible role of actin filaments. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7448–7452. [PubMed]
  • Greenwood M, Tsang A. Sequence and expression of annexin VII of Dictyostelium discoideum. Biochim Biophys Acta. 1991 Mar 26;1088(3):429–432. [PubMed]
  • Griffiths G, McDowall A, Back R, Dubochet J. On the preparation of cryosections for immunocytochemistry. J Ultrastruct Res. 1984 Oct;89(1):65–78. [PubMed]
  • Harloff C, Gerisch G, Noegel AA. Selective elimination of the contact site A protein of Dictyostelium discoideum by gene disruption. Genes Dev. 1989 Dec;3(12A):2011–2019. [PubMed]
  • Hartmann H, Noegel AA, Eckerskorn C, Rapp S, Schleicher M. Ca2+-independent F-actin capping proteins. Cap 32/34, a capping protein from Dictyostelium discoideum, does not share sequence homologies with known actin-binding proteins. J Biol Chem. 1989 Jul 25;264(21):12639–12647. [PubMed]
  • Ho WC, Allan VJ, van Meer G, Berger EG, Kreis TE. Reclustering of scattered Golgi elements occurs along microtubules. Eur J Cell Biol. 1989 Apr;48(2):250–263. [PubMed]
  • Hohmann HP, Gerisch G, Lee RW, Huttner WB. Cell-free sulfation of the contact site A glycoprotein of Dictyostelium discoideum and of a partially glycosylated precursor. J Biol Chem. 1985 Nov 5;260(25):13869–13878. [PubMed]
  • Isenberg G, Schubert P, Kreutzberg GW. Experimental approach to test the role of actin in axonal transport. Brain Res. 1980 Aug 4;194(2):588–593. [PubMed]
  • Jasmin BJ, Cartaud J, Bornens M, Changeux JP. Golgi apparatus in chick skeletal muscle: changes in its distribution during end plate development and after denervation. Proc Natl Acad Sci U S A. 1989 Sep;86(18):7218–7222. [PubMed]
  • Kelly RB. Microtubules, membrane traffic, and cell organization. Cell. 1990 Apr 6;61(1):5–7. [PubMed]
  • Kelly RB. Secretory granule and synaptic vesicle formation. Curr Opin Cell Biol. 1991 Aug;3(4):654–660. [PubMed]
  • Klausner RD, Donaldson JG, Lippincott-Schwartz J. Brefeldin A: insights into the control of membrane traffic and organelle structure. J Cell Biol. 1992 Mar;116(5):1071–1080. [PMC free article] [PubMed]
  • Kornfeld S, Mellman I. The biogenesis of lysosomes. Annu Rev Cell Biol. 1989;5:483–525. [PubMed]
  • Kreis TE. Role of microtubules in the organisation of the Golgi apparatus. Cell Motil Cytoskeleton. 1990;15(2):67–70. [PubMed]
  • Kuznetsov SA, Langford GM, Weiss DG. Actin-dependent organelle movement in squid axoplasm. Nature. 1992 Apr 23;356(6371):722–725. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Lahtinen U, Dahllöf B, Saraste J. Characterization of a 58 kDa cis-Golgi protein in pancreatic exocrine cells. J Cell Sci. 1992 Oct;103(Pt 2):321–333. [PubMed]
  • Langanger G, De Mey J, Adam H. 1,4-Diazobizyklo-[2,2,2]-Oktan (DABCO) verzögert das Ausbleichen von Immunfluoreszenzpräparaten. Mikroskopie. 1983 Sep;40(7-8):237–241. [PubMed]
  • Lippincott-Schwartz J, Donaldson JG, Schweizer A, Berger EG, Hauri HP, Yuan LC, Klausner RD. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway. Cell. 1990 Mar 9;60(5):821–836. [PubMed]
  • Loomis WF., Jr Developmental regulation of alkaline phosphatase in Dictyostelium discoideum. J Bacteriol. 1969 Oct;100(1):417–422. [PMC free article] [PubMed]
  • Loomis WF, Kuspa A. Biochemical and genetic analysis of pre-stalk specific acid phosphatase in Dictyostelium. Dev Biol. 1984 Apr;102(2):498–503. [PubMed]
  • MacLean-Fletcher SD, Pollard TD. Viscometric analysis of the gelation of Acanthamoeba extracts and purification of two gelation factors. J Cell Biol. 1980 May;85(2):414–428. [PMC free article] [PubMed]
  • Matsushima N, Creutz CE, Kretsinger RH. Polyproline, beta-turn helices. Novel secondary structures proposed for the tandem repeats within rhodopsin, synaptophysin, synexin, gliadin, RNA polymerase II, hordein, and gluten. Proteins. 1990;7(2):125–155. [PubMed]
  • McGuinness TL, Brady ST, Gruner JA, Sugimori M, Llinas R, Greengard P. Phosphorylation-dependent inhibition by synapsin I of organelle movement in squid axoplasm. J Neurosci. 1989 Dec;9(12):4138–4149. [PubMed]
  • McRobbie SJ, Newell PC. Chemoattractant-mediated changes in cytoskeletal actin of cellular slime moulds. J Cell Sci. 1984 Jun;68:139–151. [PubMed]
  • Mellman I, Simons K. The Golgi complex: in vitro veritas? Cell. 1992 Mar 6;68(5):829–840. [PubMed]
  • Nelson WJ. Regulation of cell surface polarity from bacteria to mammals. Science. 1992 Nov 6;258(5084):948–955. [PubMed]
  • Noegel AA, Gerisch G, Lottspeich F, Schleicher M. A protein with homology to the C-terminal repeat sequence of Octopus rhodopsin and synaptophysin is a member of a multigene family in Dictyostelium discoideum. FEBS Lett. 1990 Jun 18;266(1-2):118–122. [PubMed]
  • Osborn M, Weber K. Dimethylsulfoxide and the ionophore A23187 affect the arrangement of actin and induce nuclear actin paracrystals in PtK2 cells. Exp Cell Res. 1980 Sep;129(1):103–114. [PubMed]
  • Payrastre B, van Bergen en Henegouwen PM, Breton M, den Hartigh JC, Plantavid M, Verkleij AJ, Boonstra J. Phosphoinositide kinase, diacylglycerol kinase, and phospholipase C activities associated to the cytoskeleton: effect of epidermal growth factor. J Cell Biol. 1991 Oct;115(1):121–128. [PMC free article] [PubMed]
  • Pearse BM. Coated vesicles from pig brain: purification and biochemical characterization. J Mol Biol. 1975 Sep 5;97(1):93–98. [PubMed]
  • Pryer NK, Wuestehube LJ, Schekman R. Vesicle-mediated protein sorting. Annu Rev Biochem. 1992;61:471–516. [PubMed]
  • Robinson MS. 100-kD coated vesicle proteins: molecular heterogeneity and intracellular distribution studied with monoclonal antibodies. J Cell Biol. 1987 Apr;104(4):887–895. [PMC free article] [PubMed]
  • Rothberg KG, Heuser JE, Donzell WC, Ying YS, Glenney JR, Anderson RG. Caveolin, a protein component of caveolae membrane coats. Cell. 1992 Feb 21;68(4):673–682. [PubMed]
  • Schleicher M, Gerisch G, Isenberg G. New actin-binding proteins from Dictyostelium discoideum. EMBO J. 1984 Sep;3(9):2095–2100. [PubMed]
  • Schopohl D, Müller-Taubenberger A, Orthen B, Hess H, Reutter W. Purification and properties of a secreted and developmentally regulated alpha-L-fucosidase from Dictyostelium discoideum. J Biol Chem. 1992 Feb 5;267(4):2400–2405. [PubMed]
  • Serafini T, Stenbeck G, Brecht A, Lottspeich F, Orci L, Rothman JE, Wieland FT. A coat subunit of Golgi-derived non-clathrin-coated vesicles with homology to the clathrin-coated vesicle coat protein beta-adaptin. Nature. 1991 Jan 17;349(6306):215–220. [PubMed]
  • Spudich JA, Watt S. The regulation of rabbit skeletal muscle contraction. I. Biochemical studies of the interaction of the tropomyosin-troponin complex with actin and the proteolytic fragments of myosin. J Biol Chem. 1971 Aug 10;246(15):4866–4871. [PubMed]
  • Spudich JA, Kron SJ, Sheetz MP. Movement of myosin-coated beads on oriented filaments reconstituted from purified actin. Nature. 1985 Jun 13;315(6020):584–586. [PubMed]
  • Stratford CA, Brown SS. Isolation of an actin-binding protein from membranes of Dictyostelium discoideum. J Cell Biol. 1985 Mar;100(3):727–735. [PMC free article] [PubMed]
  • Swanson J. Fluorescent labeling of endocytic compartments. Methods Cell Biol. 1989;29:137–151. [PubMed]
  • Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. [PubMed]
  • Waters MG, Serafini T, Rothman JE. 'Coatomer': a cytosolic protein complex containing subunits of non-clathrin-coated Golgi transport vesicles. Nature. 1991 Jan 17;349(6306):248–251. [PubMed]
  • Weiss DG, Langford GM, Seitz-Tutter D, Keller F. Dynamic instability and motile events of native microtubules from squid axoplasm. Cell Motil Cytoskeleton. 1988;10(1-2):285–295. [PubMed]
  • Yumura S, Fukui Y. Filopodelike projections induced with dimethyl sulfoxide and their relevance to cellular polarity in Dictyostelium. J Cell Biol. 1983 Mar;96(3):857–865. [PMC free article] [PubMed]

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