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J Cell Biol. Sep 2, 1992; 118(6): 1333–1345.
PMCID: PMC2289611

Cytoplasmic dynein participates in the centrosomal localization of the Golgi complex

Abstract

The localization of the Golgi complex depends upon the integrity of the microtubule apparatus. At interphase, the Golgi has a restricted pericentriolar localization. During mitosis, it fragments into small vesicles that are dispersed throughout the cytoplasm until telophase, when they again coalesce near the centrosome. These observations have suggested that the Golgi complex utilizes a dynein-like motor to mediate its transport from the cell periphery towards the minus ends of microtubules, located at the centrosome. We utilized semi-intact cells to study the interaction of the Golgi complex with the microtubule apparatus. We show here that Golgi complexes can enter semi-intact cells and associate stably with cytoplasmic constituents. Stable association, termed here "Golgi capture," requires ATP hydrolysis and intact microtubules, and occurs maximally at physiological temperature in the presence of added cytosolic proteins. Once translocated into the semi-intact cell cytoplasm, exogenous Golgi complexes display a distribution similar to endogenous Golgi complexes, near the microtubule-organizing center. The process of Golgi capture requires cytoplasmic tubulin, and is abolished if cytoplasmic dynein is immunodepleted from the cytosol. Cytoplasmic dynein, prepared from CHO cell cytosol, restores Golgi capture activity to reactions carried out with dynein immuno-depleted cytosol. These results indicate that cytoplasmic dynein can interact with isolated Golgi complexes, and participate in their accumulation near the centrosomes of semi-intact, recipient cells. Thus, cytoplasmic dynein appears to play a role in determining the subcellular localization of the Golgi complex.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allan VJ, Vale RD. Cell cycle control of microtubule-based membrane transport and tubule formation in vitro. J Cell Biol. 1991 Apr;113(2):347–359. [PMC free article] [PubMed]
  • Balch WE, Dunphy WG, Braell WA, Rothman JE. Reconstitution of the transport of protein between successive compartments of the Golgi measured by the coupled incorporation of N-acetylglucosamine. Cell. 1984 Dec;39(2 Pt 1):405–416. [PubMed]
  • Ball EH, Singer SJ. Mitochondria are associated with microtubules and not with intermediate filaments in cultured fibroblasts. Proc Natl Acad Sci U S A. 1982 Jan;79(1):123–126. [PMC free article] [PubMed]
  • Beckers CJ, Keller DS, Balch WE. Semi-intact cells permeable to macromolecules: use in reconstitution of protein transport from the endoplasmic reticulum to the Golgi complex. Cell. 1987 Aug 14;50(4):523–534. [PubMed]
  • Bhisey AN, Freed JJ. Altered movement of endosomes in colchicine-treated cultured macrophages. Exp Cell Res. 1971 Feb;64(2):430–438. [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]
  • Braell WA, Balch WE, Dobbertin DC, Rothman JE. The glycoprotein that is transported between successive compartments of the Golgi in a cell-free system resides in stacks of cisternae. Cell. 1984 Dec;39(3 Pt 2):511–524. [PubMed]
  • Burnette WN. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. [PubMed]
  • Clary DO, Rothman JE. Purification of three related peripheral membrane proteins needed for vesicular transport. J Biol Chem. 1990 Jun 15;265(17):10109–10117. [PubMed]
  • Collot M, Louvard D, Singer SJ. Lysosomes are associated with microtubules and not with intermediate filaments in cultured fibroblasts. Proc Natl Acad Sci U S A. 1984 Feb;81(3):788–792. [PMC free article] [PubMed]
  • Cooper MS, Cornell-Bell AH, Chernjavsky A, Dani JW, Smith SJ. Tubulovesicular processes emerge from trans-Golgi cisternae, extend along microtubules, and interlink adjacent trans-golgi elements into a reticulum. Cell. 1990 Apr 6;61(1):135–145. [PubMed]
  • Deng Y, DeCourcy K, Storrie B. Intermixing of resident Golgi membrane proteins in rat-hamster polykaryons appears to depend on organelle coalescence. Eur J Cell Biol. 1992 Feb;57(1):1–11. [PubMed]
  • Donaldson JG, Lippincott-Schwartz J, Klausner RD. Guanine nucleotides modulate the effects of brefeldin A in semipermeable cells: regulation of the association of a 110-kD peripheral membrane protein with the Golgi apparatus. J Cell Biol. 1991 Feb;112(4):579–588. [PMC free article] [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]
  • Dunphy WG, Rothman JE. Compartmentation of asparagine-linked oligosaccharide processing in the Golgi apparatus. J Cell Biol. 1983 Jul;97(1):270–275. [PMC free article] [PubMed]
  • Dunphy WG, Brands R, Rothman JE. Attachment of terminal N-acetylglucosamine to asparagine-linked oligosaccharides occurs in central cisternae of the Golgi stack. Cell. 1985 Feb;40(2):463–472. [PubMed]
  • Freed JJ, Lebowitz MM. The association of a class of saltatory movements with microtubules in cultured cells. J Cell Biol. 1970 May;45(2):334–354. [PMC free article] [PubMed]
  • Gibbons IR, Lee-Eiford A, Mocz G, Phillipson CA, Tang WJ, Gibbons BH. Photosensitized cleavage of dynein heavy chains. Cleavage at the "V1 site" by irradiation at 365 nm in the presence of ATP and vanadate. J Biol Chem. 1987 Feb 25;262(6):2780–2786. [PubMed]
  • Goda Y, Pfeffer SR. Selective recycling of the mannose 6-phosphate/IGF-II receptor to the trans Golgi network in vitro. Cell. 1988 Oct 21;55(2):309–320. [PubMed]
  • Goda Y, Pfeffer SR. Identification of a novel, N-ethylmaleimide-sensitive cytosolic factor required for vesicular transport from endosomes to the trans-Golgi network in vitro. J Cell Biol. 1991 Mar;112(5):823–831. [PMC free article] [PubMed]
  • Gottlieb C, Baenziger J, Kornfeld S. Deficient uridine diphosphate-N-acetylglucosamine:glycoprotein N-acetylglucosaminyltransferase activity in a clone of Chinese hamster ovary cells with altered surface glycoproteins. J Biol Chem. 1975 May 10;250(9):3303–3309. [PubMed]
  • Herman B, Albertini DF. A time-lapse video image intensification analysis of cytoplasmic organelle movements during endosome translocation. J Cell Biol. 1984 Feb;98(2):565–576. [PMC free article] [PubMed]
  • Heuser J. Changes in lysosome shape and distribution correlated with changes in cytoplasmic pH. J Cell Biol. 1989 Mar;108(3):855–864. [PMC free article] [PubMed]
  • Hirokawa N. Cross-linker system between neurofilaments, microtubules, and membranous organelles in frog axons revealed by the quick-freeze, deep-etching method. J Cell Biol. 1982 Jul;94(1):129–142. [PMC free article] [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]
  • Hollenbeck PJ, Swanson JA. Radial extension of macrophage tubular lysosomes supported by kinesin. Nature. 1990 Aug 30;346(6287):864–866. [PubMed]
  • Karsenti E, Newport J, Hubble R, Kirschner M. Interconversion of metaphase and interphase microtubule arrays, as studied by the injection of centrosomes and nuclei into Xenopus eggs. J Cell Biol. 1984 May;98(5):1730–1745. [PMC free article] [PubMed]
  • Kreis TE. Role of microtubules in the organisation of the Golgi apparatus. Cell Motil Cytoskeleton. 1990;15(2):67–70. [PubMed]
  • Lee-Eiford A, Ow RA, Gibbons IR. Specific cleavage of dynein heavy chains by ultraviolet irradiation in the presence of ATP and vanadate. J Biol Chem. 1986 Feb 15;261(5):2337–2342. [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]
  • Lucocq JM, Pryde JG, Berger EG, Warren G. A mitotic form of the Golgi apparatus in HeLa cells. J Cell Biol. 1987 Apr;104(4):865–874. [PMC free article] [PubMed]
  • Lucocq JM, Berger EG, Warren G. Mitotic Golgi fragments in HeLa cells and their role in the reassembly pathway. J Cell Biol. 1989 Aug;109(2):463–474. [PMC free article] [PubMed]
  • Lye RJ, Porter ME, Scholey JM, McIntosh JR. Identification of a microtubule-based cytoplasmic motor in the nematode C. elegans. Cell. 1987 Oct 23;51(2):309–318. [PubMed]
  • Matteoni R, Kreis TE. Translocation and clustering of endosomes and lysosomes depends on microtubules. J Cell Biol. 1987 Sep;105(3):1253–1265. [PMC free article] [PubMed]
  • McIntosh JR, Porter ME. Enzymes for microtubule-dependent motility. J Biol Chem. 1989 Apr 15;264(11):6001–6004. [PubMed]
  • Melançon P, Glick BS, Malhotra V, Weidman PJ, Serafini T, Gleason ML, Orci L, Rothman JE. Involvement of GTP-binding "G" proteins in transport through the Golgi stack. Cell. 1987 Dec 24;51(6):1053–1062. [PubMed]
  • Parton RG, Dotti CG, Bacallao R, Kurtz I, Simons K, Prydz K. pH-induced microtubule-dependent redistribution of late endosomes in neuronal and epithelial cells. J Cell Biol. 1991 Apr;113(2):261–274. [PMC free article] [PubMed]
  • Paschal BM, Vallee RB. Retrograde transport by the microtubule-associated protein MAP 1C. Nature. 1987 Nov 12;330(6144):181–183. [PubMed]
  • Paschal BM, Shpetner HS, Vallee RB. MAP 1C is a microtubule-activated ATPase which translocates microtubules in vitro and has dynein-like properties. J Cell Biol. 1987 Sep;105(3):1273–1282. [PMC free article] [PubMed]
  • Pavelka M, Ellinger A. Effect of colchicine on the Golgi complex of rat pancreatic acinar cells. J Cell Biol. 1983 Sep;97(3):737–748. [PMC free article] [PubMed]
  • Pfarr CM, Coue M, Grissom PM, Hays TS, Porter ME, McIntosh JR. Cytoplasmic dynein is localized to kinetochores during mitosis. Nature. 1990 May 17;345(6272):263–265. [PubMed]
  • Rambourg A, Clermont Y. Three-dimensional electron microscopy: structure of the Golgi apparatus. Eur J Cell Biol. 1990 Apr;51(2):189–200. [PubMed]
  • Rebhun LI. Polarized intracellular particle transport: saltatory movements and cytoplasmic streaming. Int Rev Cytol. 1972;32:93–137. [PubMed]
  • ROBBINS E, GONATAS NK. HISTOCHEMICAL AND ULTRASTRUCTURAL STUDIES ON HELA CELL CULTURES EXPOSED TO SPINDLE INHIBITORS WITH SPECIAL REFERENCE TO THE INTERPHASE CELL. J Histochem Cytochem. 1964 Sep;12:704–711. [PubMed]
  • Rogalski AA, Bergmann JE, Singer SJ. Effect of microtubule assembly status on the intracellular processing and surface expression of an integral protein of the plasma membrane. J Cell Biol. 1984 Sep;99(3):1101–1109. [PMC free article] [PubMed]
  • Rothman JE, Orci L. Movement of proteins through the Golgi stack: a molecular dissection of vesicular transport. FASEB J. 1990 Mar;4(5):1460–1468. [PubMed]
  • Schnapp BJ, Reese TS. Dynein is the motor for retrograde axonal transport of organelles. Proc Natl Acad Sci U S A. 1989 Mar;86(5):1548–1552. [PMC free article] [PubMed]
  • Schroer TA, Sheetz MP. Functions of microtubule-based motors. Annu Rev Physiol. 1991;53:629–652. [PubMed]
  • Schroer TA, Sheetz MP. Two activators of microtubule-based vesicle transport. J Cell Biol. 1991 Dec;115(5):1309–1318. [PMC free article] [PubMed]
  • Schroer TA, Steuer ER, Sheetz MP. Cytoplasmic dynein is a minus end-directed motor for membranous organelles. Cell. 1989 Mar 24;56(6):937–946. [PubMed]
  • Shpetner HS, Paschal BM, Vallee RB. Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP 1C). J Cell Biol. 1988 Sep;107(3):1001–1009. [PMC free article] [PubMed]
  • Skoufias DA, Burgess TL, Wilson L. Spatial and temporal colocalization of the Golgi apparatus and microtubules rich in detyrosinated tubulin. J Cell Biol. 1990 Nov;111(5 Pt 1):1929–1937. [PMC free article] [PubMed]
  • Steuer ER, Wordeman L, Schroer TA, Sheetz MP. Localization of cytoplasmic dynein to mitotic spindles and kinetochores. Nature. 1990 May 17;345(6272):266–268. [PubMed]
  • Swanson J, Bushnell A, Silverstein SC. Tubular lysosome morphology and distribution within macrophages depend on the integrity of cytoplasmic microtubules. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1921–1925. [PMC free article] [PubMed]
  • Tabas I, Kornfeld S. Purification and characterization of a rat liver Golgi alpha-mannosidase capable of processing asparagine-linked oligosaccharides. J Biol Chem. 1979 Nov 25;254(22):11655–11663. [PubMed]
  • Terasaki M, Chen LB, Fujiwara K. Microtubules and the endoplasmic reticulum are highly interdependent structures. J Cell Biol. 1986 Oct;103(4):1557–1568. [PMC free article] [PubMed]
  • Thyberg J, Moskalewski S. Microtubules and the organization of the Golgi complex. Exp Cell Res. 1985 Jul;159(1):1–16. [PubMed]
  • Thyberg J, Moskalewski S. Subpopulations of microtubules with differential sensitivity to nocodazole: role in the structural organization of the Golgi complex and the lysosomal system. J Submicrosc Cytol Pathol. 1989 Apr;21(2):259–274. [PubMed]
  • Turner JR, Tartakoff AM. The response of the Golgi complex to microtubule alterations: the roles of metabolic energy and membrane traffic in Golgi complex organization. J Cell Biol. 1989 Nov;109(5):2081–2088. [PMC free article] [PubMed]
  • Vale RD. Microtubule-based motor proteins. Curr Opin Cell Biol. 1990 Feb;2(1):15–22. [PubMed]
  • Vale RD, Goldstein LS. One motor, many tails: an expanding repertoire of force-generating enzymes. Cell. 1990 Mar 23;60(6):883–885. [PubMed]
  • Vallee R. Cytoplasmic dynein: advances in microtubule-based motility. Trends Cell Biol. 1991 Jul;1(1):25–29. [PubMed]
  • Warren G, Davoust J, Cockcroft A. Recycling of transferrin receptors in A431 cells is inhibited during mitosis. EMBO J. 1984 Oct;3(10):2217–2225. [PMC free article] [PubMed]

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