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. 1989 Jul 1;109(1):163–177. doi: 10.1083/jcb.109.1.163

Regulation of desmosome assembly in epithelial cells: kinetics of synthesis, transport, and stabilization of desmoglein I, a major protein of the membrane core domain

PMCID: PMC2115471  PMID: 2501314

Abstract

Desmosomes are composed of two morphologically and biochemically distinct domains, a cytoplasmic plaque and membrane core. We have initiated a study of the synthesis and assembly of these domains in Madin-Darby canine kidney (MDCK) epithelial cells to understand the mechanisms involved in the formation of desmosomes. Previously, we reported the kinetics of assembly of two components of the cytoplasmic plaque domain, Desmoplakin I/II (Pasdar, M., and W. J. Nelson. 1988. J. Cell Biol. 106:677-685 and 106:687-699. We have now extended this analysis to include a major glycoprotein component of the membrane core domain, Desmoglein I (DGI; Mr = 150,000). Using metabolic labeling and inhibitors of glycoprotein processing and intracellular transport, we show that DGI biosynthesis is a sequential process with defined stages. In the absence of cell-cell contact, DGI enters a Triton X-100 soluble pool and is core glycosylated. The soluble DGI is then transported to the Golgi complex where it is first complex glycosylated and then titrated into an insoluble pool. The insoluble pool of DGI is subsequently transported to the plasma membrane and is degraded rapidly (t1/2 less than 4 h). Although this biosynthetic pathway occurs independently of cell-cell contact, induction of cell-cell contact results in dramatic increases in the efficiency and rate of titration of DGI from the soluble to the insoluble pool, and its transport to the plasma membrane where DGI becomes metabolically stable (t1/2 greater than 24 h). Taken together with our previous study of DPI/II, we conclude that newly synthesized components of the cytoplasmic plaque and membrane core domains are processed and assembled with different kinetics indicating that, at least initially, each domain is assembled separately in the cell. However, upon induction of cell-cell contact there is a rapid titration of both components into an insoluble and metabolically stable pool at the plasma membrane that is concurrent with desmosome assembly.

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Selected References

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  1. Bourguignon L. Y., Singer S. J. Transmembrane interactions and the mechanism of capping of surface receptors by their specific ligands. Proc Natl Acad Sci U S A. 1977 Nov;74(11):5031–5035. doi: 10.1073/pnas.74.11.5031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Cohen S. M., Gorbsky G., Steinberg M. S. Immunochemical characterization of related families of glycoproteins in desmosomes. J Biol Chem. 1983 Feb 25;258(4):2621–2627. [PubMed] [Google Scholar]
  3. Cowin P., Garrod D. R. Antibodies to epithelial desmosomes show wide tissue and species cross-reactivity. Nature. 1983 Mar 10;302(5904):148–150. doi: 10.1038/302148a0. [DOI] [PubMed] [Google Scholar]
  4. Cowin P., Kapprell H. P., Franke W. W., Tamkun J., Hynes R. O. Plakoglobin: a protein common to different kinds of intercellular adhering junctions. Cell. 1986 Sep 26;46(7):1063–1073. doi: 10.1016/0092-8674(86)90706-3. [DOI] [PubMed] [Google Scholar]
  5. Cowin P., Kapprell H. P., Franke W. W. The complement of desmosomal plaque proteins in different cell types. J Cell Biol. 1985 Oct;101(4):1442–1454. doi: 10.1083/jcb.101.4.1442. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cowin P., Mattey D., Garrod D. Distribution of desmosomal components in the tissues of vertebrates, studied by fluorescent antibody staining. J Cell Sci. 1984 Mar;66:119–132. doi: 10.1242/jcs.66.1.119. [DOI] [PubMed] [Google Scholar]
  7. Cowin P., Mattey D., Garrod D. Identification of desmosomal surface components (desmocollins) and inhibition of desmosome formation by specific Fab'. J Cell Sci. 1984 Aug;70:41–60. doi: 10.1242/jcs.70.1.41. [DOI] [PubMed] [Google Scholar]
  8. Czichi U., Lennarz W. J. Localization of the enzyme system for glycosylation of proteins via the lipid-linked pathway in rough endoplasmic reticulum. J Biol Chem. 1977 Nov 25;252(22):7901–7904. [PubMed] [Google Scholar]
  9. Drochmans P., Freudenstein C., Wanson J. C., Laurent L., Keenan T. W., Stadler J., Leloup R., Franke W. W. Structure and biochemical composition of desmosomes and tonofilaments isolated from calf muzzle epidermis. J Cell Biol. 1978 Nov;79(2 Pt 1):427–443. doi: 10.1083/jcb.79.2.427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dunphy W. G., Brands R., Rothman J. E. Attachment of terminal N-acetylglucosamine to asparagine-linked oligosaccharides occurs in central cisternae of the Golgi stack. Cell. 1985 Feb;40(2):463–472. doi: 10.1016/0092-8674(85)90161-8. [DOI] [PubMed] [Google Scholar]
  11. Franke W. W., Mueller H., Mittnacht S., Kapprell H. P., Jorcano J. L. Significance of two desmosome plaque-associated polypeptides of molecular weights 75 000 and 83 000. EMBO J. 1983;2(12):2211–2215. doi: 10.1002/j.1460-2075.1983.tb01725.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Franke W. W., Schmid E., Grund C., Müller H., Engelbrecht I., Moll R., Stadler J., Jarasch E. D. Antibodies to high molecular weight polypeptides of desmosomes: specific localization of a class of junctional proteins in cells and tissue. Differentiation. 1981;20(3):217–241. doi: 10.1111/j.1432-0436.1981.tb01178.x. [DOI] [PubMed] [Google Scholar]
  13. Garrod D. R. Formation of desmosomes in polarized and non-polarized epithelial cells: implications for epithelial morphogenesis. Biochem Soc Trans. 1986 Feb;14(1):172–175. doi: 10.1042/bst0140172. [DOI] [PubMed] [Google Scholar]
  14. Geiger B., Schmid E., Franke W. W. Spatial distribution of proteins specific for desmosomes and adhaerens junctions in epithelial cells demonstrated by double immunofluorescence microscopy. Differentiation. 1983;23(3):189–205. doi: 10.1111/j.1432-0436.1982.tb01283.x. [DOI] [PubMed] [Google Scholar]
  15. Gumbiner B., Stevenson B., Grimaldi A. The role of the cell adhesion molecule uvomorulin in the formation and maintenance of the epithelial junctional complex. J Cell Biol. 1988 Oct;107(4):1575–1587. doi: 10.1083/jcb.107.4.1575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hubbard S. C., Ivatt R. J. Synthesis and processing of asparagine-linked oligosaccharides. Annu Rev Biochem. 1981;50:555–583. doi: 10.1146/annurev.bi.50.070181.003011. [DOI] [PubMed] [Google Scholar]
  17. Johnson D. C., Spear P. G. Monensin inhibits the processing of herpes simplex virus glycoproteins, their transport to the cell surface, and the egress of virions from infected cells. J Virol. 1982 Sep;43(3):1102–1112. doi: 10.1128/jvi.43.3.1102-1112.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kapprell H. P., Cowin P., Franke W. W., Ponstingl H., Opferkuch H. J. Biochemical characterization of desmosomal proteins isolated from bovine muzzle epidermis: amino acid and carbohydrate composition. Eur J Cell Biol. 1985 Mar;36(2):217–229. [PubMed] [Google Scholar]
  19. Kelly D. E. Fine structure of desmosomes. , hemidesmosomes, and an adepidermal globular layer in developing newt epidermis. J Cell Biol. 1966 Jan;28(1):51–72. doi: 10.1083/jcb.28.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kelly D. E., Shienvold F. L. The desmosome: fine structural studies with freeze-fracture replication and tannic acid staining of sectioned epidermis. Cell Tissue Res. 1976 Sep 20;172(3):309–323. doi: 10.1007/BF00399514. [DOI] [PubMed] [Google Scholar]
  21. Matlin K. S., Simons K. Reduced temperature prevents transfer of a membrane glycoprotein to the cell surface but does not prevent terminal glycosylation. Cell. 1983 Aug;34(1):233–243. doi: 10.1016/0092-8674(83)90154-x. [DOI] [PubMed] [Google Scholar]
  22. Mattey D. L., Garrod D. R. Calcium-induced desmosome formation in cultured kidney epithelial cells. J Cell Sci. 1986 Sep;85:95–111. doi: 10.1242/jcs.85.1.95. [DOI] [PubMed] [Google Scholar]
  23. Miller K., Mattey D., Measures H., Hopkins C., Garrod D. Localisation of the protein and glycoprotein components of bovine nasal epithelial desmosomes by immunoelectron microscopy. EMBO J. 1987 Apr;6(4):885–889. doi: 10.1002/j.1460-2075.1987.tb04834.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mueller H., Franke W. W. Biochemical and immunological characterization of desmoplakins I and II, the major polypeptides of the desmosomal plaque. J Mol Biol. 1983 Feb 5;163(4):647–671. doi: 10.1016/0022-2836(83)90116-x. [DOI] [PubMed] [Google Scholar]
  25. Nagafuchi A., Shirayoshi Y., Okazaki K., Yasuda K., Takeichi M. Transformation of cell adhesion properties by exogenously introduced E-cadherin cDNA. Nature. 1987 Sep 24;329(6137):341–343. doi: 10.1038/329341a0. [DOI] [PubMed] [Google Scholar]
  26. Nelson W. J., Colaço C. A., Lazarides E. Involvement of spectrin in cell-surface receptor capping in lymphocytes. Proc Natl Acad Sci U S A. 1983 Mar;80(6):1626–1630. doi: 10.1073/pnas.80.6.1626. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Nelson W. J., Veshnock P. J. Dynamics of membrane-skeleton (fodrin) organization during development of polarity in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1986 Nov;103(5):1751–1765. doi: 10.1083/jcb.103.5.1751. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Nelson W. J., Veshnock P. J. Modulation of fodrin (membrane skeleton) stability by cell-cell contact in Madin-Darby canine kidney epithelial cells. J Cell Biol. 1987 Jun;104(6):1527–1537. doi: 10.1083/jcb.104.6.1527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. OVERTON J. Desmosome development in normal and reassociating cells in the early chick blastoderm. Dev Biol. 1962 Jun;4:532–548. doi: 10.1016/0012-1606(62)90056-8. [DOI] [PubMed] [Google Scholar]
  30. Overton J. Experimental manipulation of desmosome formation. J Cell Biol. 1973 Mar;56(3):636–646. doi: 10.1083/jcb.56.3.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Overton J. Experiments with junctions of the adhaerens type. Curr Top Dev Biol. 1975;10:1–34. doi: 10.1016/s0070-2153(08)60037-x. [DOI] [PubMed] [Google Scholar]
  32. Overton J. Inhibition of desmosome formation with tunicamycin and with lectin in corneal cell aggregates. Dev Biol. 1982 Jul;92(1):66–72. doi: 10.1016/0012-1606(82)90151-8. [DOI] [PubMed] [Google Scholar]
  33. Overton J. Selective formation of desmosomes in chick cell reaggregates. Dev Biol. 1974 Aug;39(2):210–225. doi: 10.1016/0012-1606(74)90236-x. [DOI] [PubMed] [Google Scholar]
  34. Pasdar M., Nelson W. J. Kinetics of desmosome assembly in Madin-Darby canine kidney epithelial cells: temporal and spatial regulation of desmoplakin organization and stabilization upon cell-cell contact. I. Biochemical analysis. J Cell Biol. 1988 Mar;106(3):677–685. doi: 10.1083/jcb.106.3.677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Pasdar M., Nelson W. J. Kinetics of desmosome assembly in Madin-Darby canine kidney epithelial cells: temporal and spatial regulation of desmoplakin organization and stabilization upon cell-cell contact. II. Morphological analysis. J Cell Biol. 1988 Mar;106(3):687–695. doi: 10.1083/jcb.106.3.687. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Penn E. J., Burdett I. D., Hobson C., Magee A. I., Rees D. A. Structure and assembly of desmosome junctions: biosynthesis and turnover of the major desmosome components of Madin-Darby canine kidney cells in low calcium medium. J Cell Biol. 1987 Nov;105(5):2327–2334. doi: 10.1083/jcb.105.5.2327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Penn E. J., Hobson C., Rees D. A., Magee A. I. Structure and assembly of desmosome junctions: biosynthesis, processing, and transport of the major protein and glycoprotein components in cultured epithelial cells. J Cell Biol. 1987 Jul;105(1):57–68. doi: 10.1083/jcb.105.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. RODRIGUEZ J., DEINHARDT F. Preparation of a semipermanent mounting medium for fluorescent antibody studies. Virology. 1960 Oct;12:316–317. doi: 10.1016/0042-6822(60)90205-1. [DOI] [PubMed] [Google Scholar]
  39. Skerrow C. J., Hunter I., Skerrow D. Dissection of the bovine epidermal desmosome into cytoplasmic protein and membrane glycoprotein domains. J Cell Sci. 1987 Apr;87(Pt 3):411–421. doi: 10.1242/jcs.87.3.411. [DOI] [PubMed] [Google Scholar]
  40. Skerrow C. J., Matoltsy A. G. Isolation of epidermal desmosomes. J Cell Biol. 1974 Nov;63(2 Pt 1):515–523. doi: 10.1083/jcb.63.2.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Suhrbier A., Garrod D. An investigation of the molecular components of desmosomes in epithelial cells of five vertebrates. J Cell Sci. 1986 Mar;81:223–242. doi: 10.1242/jcs.81.1.223. [DOI] [PubMed] [Google Scholar]
  42. Tarentino A. L., Trimble R. B., Maley F. endo-beta-N-Acetylglucosaminidase from Streptomyces plicatus. Methods Enzymol. 1978;50:574–580. doi: 10.1016/0076-6879(78)50065-7. [DOI] [PubMed] [Google Scholar]
  43. Tartakoff A. M. Perturbation of vesicular traffic with the carboxylic ionophore monensin. Cell. 1983 Apr;32(4):1026–1028. doi: 10.1016/0092-8674(83)90286-6. [DOI] [PubMed] [Google Scholar]
  44. Tartakoff A. M., Vassalli P. Plasma cell immunoglobulin secretion: arrest is accompanied by alterations of the golgi complex. J Exp Med. 1977 Nov 1;146(5):1332–1345. doi: 10.1084/jem.146.5.1332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tkacz J. S., Lampen O. Tunicamycin inhibition of polyisoprenyl N-acetylglucosaminyl pyrophosphate formation in calf-liver microsomes. Biochem Biophys Res Commun. 1975 Jul 8;65(1):248–257. doi: 10.1016/s0006-291x(75)80086-6. [DOI] [PubMed] [Google Scholar]
  46. 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. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

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