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. 1995 Apr 1;129(1):203–217. doi: 10.1083/jcb.129.1.203

The molecular organization of endothelial cell to cell junctions: differential association of plakoglobin, beta-catenin, and alpha- catenin with vascular endothelial cadherin (VE-cadherin)

PMCID: PMC2120375  PMID: 7698986

Abstract

In this paper we report that the assembly of interendothelial junctions containing the cell type-specific vascular endothelial cadherin (VE- cadherin or cadherin-5) is a dynamic process which is affected by the functional state of the cells. Immunofluorescence double labeling of endothelial cells (EC) cultures indicated that VE-cadherin, alpha- catenin, and beta-catenin colocalized in areas of cell to cell contact both in sparse and confluent EC monolayers. In contrast, plakoglobin became associated with cell-cell junctions only in tightly confluent cells concomitantly with an increase in its protein and mRNA levels. Furthermore, the amount of plakoglobin coimmunoprecipitated with VE- cadherin, increased in closely packed monolayers. Artificial wounding of confluent EC monolayers resulted in a major reorganization of VE- cadherin, alpha-catenin, beta-catenin, and plakoglobin. All these proteins decreased in intensity at the boundaries of EC migrating into the lesion. In contrast, EC located immediately behind the migrating front retained junctional VE-cadherin, alpha-catenin, and beta-catenin while plakoglobin was absent from these sites. In line with this observation, the amount of plakoglobin coimmunoprecipitated with VE- cadherin decreased in migrating EC. These data suggest that VE- cadherin, alpha-catenin, and beta-catenin are already associated with each other at early stages of intercellular adhesion and become readily organized at nascant cell contacts. Plakoglobin, on the other hand, associates with junctions only when cells approach confluence. When cells migrate, this order is reversed, namely, plakoglobin dissociates first and, then, VE-cadherin, alpha-catenin, and beta-catenin disassemble from the junctions. The late association of plakoglobin with junctions suggests that while VE-cadherin/alpha-catenin/beta- catenin complex can function as an early recognition mechanism between EC, the formation of mature, cytoskeleton-bound junctions requires plakoglobin synthesis and organization.

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Selected References

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  1. Alexander J. S., Blaschuk O. W., Haselton F. R. An N-cadherin-like protein contributes to solute barrier maintenance in cultured endothelium. J Cell Physiol. 1993 Sep;156(3):610–618. doi: 10.1002/jcp.1041560321. [DOI] [PubMed] [Google Scholar]
  2. Ausio J. Structure and dynamics of transcriptionally active chromatin. J Cell Sci. 1992 May;102(Pt 1):1–5. doi: 10.1242/jcs.102.1.1. [DOI] [PubMed] [Google Scholar]
  3. Ayalon O., Sabanai H., Lampugnani M. G., Dejana E., Geiger B. Spatial and temporal relationships between cadherins and PECAM-1 in cell-cell junctions of human endothelial cells. J Cell Biol. 1994 Jul;126(1):247–258. doi: 10.1083/jcb.126.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradley R. S., Cowin P., Brown A. M. Expression of Wnt-1 in PC12 cells results in modulation of plakoglobin and E-cadherin and increased cellular adhesion. J Cell Biol. 1993 Dec;123(6 Pt 2):1857–1865. doi: 10.1083/jcb.123.6.1857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Citi S. Protein kinase inhibitors prevent junction dissociation induced by low extracellular calcium in MDCK epithelial cells. J Cell Biol. 1992 Apr;117(1):169–178. doi: 10.1083/jcb.117.1.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Engerman R. L., Pfaffenbach D., Davis M. D. Cell turnover of capillaries. Lab Invest. 1967 Dec;17(6):738–743. [PubMed] [Google Scholar]
  8. Folkman J., Shing Y. Angiogenesis. J Biol Chem. 1992 Jun 5;267(16):10931–10934. [PubMed] [Google Scholar]
  9. Franke W. W., Goldschmidt M. D., Zimbelmann R., Mueller H. M., Schiller D. L., Cowin P. Molecular cloning and amino acid sequence of human plakoglobin, the common junctional plaque protein. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4027–4031. doi: 10.1073/pnas.86.11.4027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Franke W. W., Kapprell H. P., Cowin P. Immunolocalization of plakoglobin in endothelial junctions: identification as a special type of Zonulae adhaerentes. Biol Cell. 1987;59(3):205–218. doi: 10.1111/j.1768-322x.1987.tb00532.x. [DOI] [PubMed] [Google Scholar]
  11. Fujimori T., Takeichi M. Disruption of epithelial cell-cell adhesion by exogenous expression of a mutated nonfunctional N-cadherin. Mol Biol Cell. 1993 Jan;4(1):37–47. doi: 10.1091/mbc.4.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Geiger B., Ginsberg D. The cytoplasmic domain of adherens-type junctions. Cell Motil Cytoskeleton. 1991;20(1):1–6. doi: 10.1002/cm.970200102. [DOI] [PubMed] [Google Scholar]
  13. Geiger B., Volberg T., Ginsberg D., Bitzur S., Sabanay I., Hynes R. O. Broad spectrum pan-cadherin antibodies, reactive with the C-terminal 24 amino acid residues of N-cadherin. J Cell Sci. 1990 Dec;97(Pt 4):607–614. doi: 10.1242/jcs.97.4.607. [DOI] [PubMed] [Google Scholar]
  14. Golay J., Capucci A., Arsura M., Castellano M., Rizzo V., Introna M. Expression of c-myb and B-myb, but not A-myb, correlates with proliferation in human hematopoietic cells. Blood. 1991 Jan 1;77(1):149–158. [PubMed] [Google Scholar]
  15. Grunwald G. B. The structural and functional analysis of cadherin calcium-dependent cell adhesion molecules. Curr Opin Cell Biol. 1993 Oct;5(5):797–805. doi: 10.1016/0955-0674(93)90028-o. [DOI] [PubMed] [Google Scholar]
  16. Hinck L., Nelson W. J., Papkoff J. Wnt-1 modulates cell-cell adhesion in mammalian cells by stabilizing beta-catenin binding to the cell adhesion protein cadherin. J Cell Biol. 1994 Mar;124(5):729–741. doi: 10.1083/jcb.124.5.729. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hinck L., Näthke I. S., Papkoff J., Nelson W. J. Dynamics of cadherin/catenin complex formation: novel protein interactions and pathways of complex assembly. J Cell Biol. 1994 Jun;125(6):1327–1340. doi: 10.1083/jcb.125.6.1327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hirano S., Kimoto N., Shimoyama Y., Hirohashi S., Takeichi M. Identification of a neural alpha-catenin as a key regulator of cadherin function and multicellular organization. Cell. 1992 Jul 24;70(2):293–301. doi: 10.1016/0092-8674(92)90103-j. [DOI] [PubMed] [Google Scholar]
  19. Hirano S., Nose A., Hatta K., Kawakami A., Takeichi M. Calcium-dependent cell-cell adhesion molecules (cadherins): subclass specificities and possible involvement of actin bundles. J Cell Biol. 1987 Dec;105(6 Pt 1):2501–2510. doi: 10.1083/jcb.105.6.2501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Jongen W. M., Fitzgerald D. J., Asamoto M., Piccoli C., Slaga T. J., Gros D., Takeichi M., Yamasaki H. Regulation of connexin 43-mediated gap junctional intercellular communication by Ca2+ in mouse epidermal cells is controlled by E-cadherin. J Cell Biol. 1991 Aug;114(3):545–555. doi: 10.1083/jcb.114.3.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kartenbeck J., Schmelz M., Franke W. W., Geiger B. Endocytosis of junctional cadherins in bovine kidney epithelial (MDBK) cells cultured in low Ca2+ ion medium. J Cell Biol. 1991 May;113(4):881–892. doi: 10.1083/jcb.113.4.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kemler R. From cadherins to catenins: cytoplasmic protein interactions and regulation of cell adhesion. Trends Genet. 1993 Sep;9(9):317–321. doi: 10.1016/0168-9525(93)90250-l. [DOI] [PubMed] [Google Scholar]
  23. Kintner C. Regulation of embryonic cell adhesion by the cadherin cytoplasmic domain. Cell. 1992 Apr 17;69(2):225–236. doi: 10.1016/0092-8674(92)90404-z. [DOI] [PubMed] [Google Scholar]
  24. Knudsen K. A., Wheelock M. J. Plakoglobin, or an 83-kD homologue distinct from beta-catenin, interacts with E-cadherin and N-cadherin. J Cell Biol. 1992 Aug;118(3):671–679. doi: 10.1083/jcb.118.3.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lampugnani M. G., Resnati M., Raiteri M., Pigott R., Pisacane A., Houen G., Ruco L. P., Dejana E. A novel endothelial-specific membrane protein is a marker of cell-cell contacts. J Cell Biol. 1992 Sep;118(6):1511–1522. doi: 10.1083/jcb.118.6.1511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Leach L., Clark P., Lampugnani M. G., Arroyo A. G., Dejana E., Firth J. A. Immunoelectron characterisation of the inter-endothelial junctions of human term placenta. J Cell Sci. 1993 Apr;104(Pt 4):1073–1081. doi: 10.1242/jcs.104.4.1073. [DOI] [PubMed] [Google Scholar]
  27. Liaw C. W., Cannon C., Power M. D., Kiboneka P. K., Rubin L. L. Identification and cloning of two species of cadherins in bovine endothelial cells. EMBO J. 1990 Sep;9(9):2701–2708. doi: 10.1002/j.1460-2075.1990.tb07456.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McCrea P. D., Gumbiner B. M. Purification of a 92-kDa cytoplasmic protein tightly associated with the cell-cell adhesion molecule E-cadherin (uvomorulin). Characterization and extractability of the protein complex from the cell cytostructure. J Biol Chem. 1991 Mar 5;266(7):4514–4520. [PubMed] [Google Scholar]
  29. McCrea P. D., Turck C. W., Gumbiner B. A homolog of the armadillo protein in Drosophila (plakoglobin) associated with E-cadherin. Science. 1991 Nov 29;254(5036):1359–1361. doi: 10.1126/science.1962194. [DOI] [PubMed] [Google Scholar]
  30. McNeill H., Ozawa M., Kemler R., Nelson W. J. Novel function of the cell adhesion molecule uvomorulin as an inducer of cell surface polarity. Cell. 1990 Jul 27;62(2):309–316. doi: 10.1016/0092-8674(90)90368-o. [DOI] [PubMed] [Google Scholar]
  31. McNeill H., Ryan T. A., Smith S. J., Nelson W. J. Spatial and temporal dissection of immediate and early events following cadherin-mediated epithelial cell adhesion. J Cell Biol. 1993 Mar;120(5):1217–1226. doi: 10.1083/jcb.120.5.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Meyer R. A., Laird D. W., Revel J. P., Johnson R. G. Inhibition of gap junction and adherens junction assembly by connexin and A-CAM antibodies. J Cell Biol. 1992 Oct;119(1):179–189. doi: 10.1083/jcb.119.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Muller W. A., Gimbrone M. A., Jr Plasmalemmal proteins of cultured vascular endothelial cells exhibit apical-basal polarity: analysis by surface-selective iodination. J Cell Biol. 1986 Dec;103(6 Pt 1):2389–2402. doi: 10.1083/jcb.103.6.2389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Nagafuchi A., Takeichi M. Cell binding function of E-cadherin is regulated by the cytoplasmic domain. EMBO J. 1988 Dec 1;7(12):3679–3684. doi: 10.1002/j.1460-2075.1988.tb03249.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Nagafuchi A., Takeichi M. Transmembrane control of cadherin-mediated cell adhesion: a 94 kDa protein functionally associated with a specific region of the cytoplasmic domain of E-cadherin. Cell Regul. 1989 Nov;1(1):37–44. doi: 10.1091/mbc.1.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Newman P. J., Berndt M. C., Gorski J., White G. C., 2nd, Lyman S., Paddock C., Muller W. A. PECAM-1 (CD31) cloning and relation to adhesion molecules of the immunoglobulin gene superfamily. Science. 1990 Mar 9;247(4947):1219–1222. doi: 10.1126/science.1690453. [DOI] [PubMed] [Google Scholar]
  37. Näthke I. S., Hinck L., Swedlow J. R., Papkoff J., Nelson W. J. Defining interactions and distributions of cadherin and catenin complexes in polarized epithelial cells. J Cell Biol. 1994 Jun;125(6):1341–1352. doi: 10.1083/jcb.125.6.1341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Ozawa M., Baribault H., Kemler R. The cytoplasmic domain of the cell adhesion molecule uvomorulin associates with three independent proteins structurally related in different species. EMBO J. 1989 Jun;8(6):1711–1717. doi: 10.1002/j.1460-2075.1989.tb03563.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Ozawa M., Engel J., Kemler R. Single amino acid substitutions in one Ca2+ binding site of uvomorulin abolish the adhesive function. Cell. 1990 Nov 30;63(5):1033–1038. doi: 10.1016/0092-8674(90)90506-a. [DOI] [PubMed] [Google Scholar]
  40. Ozawa M., Kemler R. Molecular organization of the uvomorulin-catenin complex. J Cell Biol. 1992 Feb;116(4):989–996. doi: 10.1083/jcb.116.4.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Ozawa M., Ringwald M., Kemler R. Uvomorulin-catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4246–4250. doi: 10.1073/pnas.87.11.4246. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Pepper M. S., Montesano R., el Aoumari A., Gros D., Orci L., Meda P. Coupling and connexin 43 expression in microvascular and large vessel endothelial cells. Am J Physiol. 1992 May;262(5 Pt 1):C1246–C1257. doi: 10.1152/ajpcell.1992.262.5.C1246. [DOI] [PubMed] [Google Scholar]
  43. Pepper M. S., Sappino A. P., Montesano R., Orci L., Vassalli J. D. Plasminogen activator inhibitor-1 is induced in migrating endothelial cells. J Cell Physiol. 1992 Oct;153(1):129–139. doi: 10.1002/jcp.1041530117. [DOI] [PubMed] [Google Scholar]
  44. Piepenhagen P. A., Nelson W. J. Defining E-cadherin-associated protein complexes in epithelial cells: plakoglobin, beta- and gamma-catenin are distinct components. J Cell Sci. 1993 Mar;104(Pt 3):751–762. doi: 10.1242/jcs.104.3.751. [DOI] [PubMed] [Google Scholar]
  45. Rubin L. L., Hall D. E., Porter S., Barbu K., Cannon C., Horner H. C., Janatpour M., Liaw C. W., Manning K., Morales J. A cell culture model of the blood-brain barrier. J Cell Biol. 1991 Dec;115(6):1725–1735. doi: 10.1083/jcb.115.6.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Schmelz M., Franke W. W. Complexus adhaerentes, a new group of desmoplakin-containing junctions in endothelial cells: the syndesmos connecting retothelial cells of lymph nodes. Eur J Cell Biol. 1993 Aug;61(2):274–289. [PubMed] [Google Scholar]
  47. Schwartz S. M., Haudenschild C. C., Eddy E. M. Endothelial regneration. I. Quantitative analysis of initial stages of endothelial regeneration in rat aortic intima. Lab Invest. 1978 May;38(5):568–580. [PubMed] [Google Scholar]
  48. Shasby D. M., Shasby S. S. Effects of calcium on transendothelial albumin transfer and electrical resistance. J Appl Physiol (1985) 1986 Jan;60(1):71–79. doi: 10.1152/jappl.1986.60.1.71. [DOI] [PubMed] [Google Scholar]
  49. Shimoyama Y., Nagafuchi A., Fujita S., Gotoh M., Takeichi M., Tsukita S., Hirohashi S. Cadherin dysfunction in a human cancer cell line: possible involvement of loss of alpha-catenin expression in reduced cell-cell adhesiveness. Cancer Res. 1992 Oct 15;52(20):5770–5774. [PubMed] [Google Scholar]
  50. Sholley M. M., Gimbrone M. A., Jr, Cotran R. S. Cellular migration and replication in endothelial regeneration: a study using irradiated endothelial cultures. Lab Invest. 1977 Jan;36(1):18–25. [PubMed] [Google Scholar]
  51. Shore E. M., Nelson W. J. Biosynthesis of the cell adhesion molecule uvomorulin (E-cadherin) in Madin-Darby canine kidney epithelial cells. J Biol Chem. 1991 Oct 15;266(29):19672–19680. [PubMed] [Google Scholar]
  52. Simionescu M., Simionescu N. Endothelial transport of macromolecules: transcytosis and endocytosis. A look from cell biology. Cell Biol Rev. 1991;25(1):5–78. [PubMed] [Google Scholar]
  53. Simmons D. L., Walker C., Power C., Pigott R. Molecular cloning of CD31, a putative intercellular adhesion molecule closely related to carcinoembryonic antigen. J Exp Med. 1990 Jun 1;171(6):2147–2152. doi: 10.1084/jem.171.6.2147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Stolz D. B., Bannish G., Jacobson B. S. The role of the cytoskeleton and intercellular junctions in the transcellular membrane protein polarity of bovine aortic endothelial cells in vitro. J Cell Sci. 1992 Sep;103(Pt 1):53–68. doi: 10.1242/jcs.103.1.53. [DOI] [PubMed] [Google Scholar]
  55. Suzuki S., Sano K., Tanihara H. Diversity of the cadherin family: evidence for eight new cadherins in nervous tissue. Cell Regul. 1991 Apr;2(4):261–270. doi: 10.1091/mbc.2.4.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Tabor S., Richardson C. C. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. doi: 10.1073/pnas.84.14.4767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  57. Tsukita S., Tsukita S., Nagafuchi A., Yonemura S. Molecular linkage between cadherins and actin filaments in cell-cell adherens junctions. Curr Opin Cell Biol. 1992 Oct;4(5):834–839. doi: 10.1016/0955-0674(92)90108-o. [DOI] [PubMed] [Google Scholar]
  58. Volberg T., Geiger B., Kartenbeck J., Franke W. W. Changes in membrane-microfilament interaction in intercellular adherens junctions upon removal of extracellular Ca2+ ions. J Cell Biol. 1986 May;102(5):1832–1842. doi: 10.1083/jcb.102.5.1832. [DOI] [PMC free article] [PubMed] [Google Scholar]

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