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. 1998 Apr 1;17(7):1919–1929. doi: 10.1093/emboj/17.7.1919

Carbohydrate-mediated Golgi to cell surface transport and apical targeting of membrane proteins.

A Gut 1, F Kappeler 1, N Hyka 1, M S Balda 1, H P Hauri 1, K Matter 1
PMCID: PMC1170538  PMID: 9524115

Abstract

Polarized expression of most epithelial plasma membrane proteins is achieved by selective transport from the Golgi apparatus or from endosomes to a specific cell surface domain. In Madin-Darby canine kidney (MDCK) cells, basolateral sorting generally depends on distinct cytoplasmic targeting determinants. Inactivation of these signals often resulted in apical expression, suggesting that apical transport of transmembrane proteins occurs either by default or is mediated by widely distributed characteristics of membrane glycoproteins. We tested the hypothesis of N-linked carbohydrates acting as apical targeting signals using three different membrane proteins. The first two are normally not glycosylated and the third one is a glycoprotein. In all three cases, N-linked carbohydrates were clearly able to mediate apical targeting and transport. Cell surface transport of proteins containing cytoplasmic basolateral targeting determinants was not significantly affected by N-linked sugars. In the absence of glycosylation and a basolateral sorting signal, the reporter proteins accumulated in the Golgi complex of MDCK as well as CHO cells, indicating that efficient transport from the Golgi apparatus to the cell surface is signal-mediated in polarized and non-polarized cells.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Balda M. S., Whitney J. A., Flores C., González S., Cereijido M., Matter K. Functional dissociation of paracellular permeability and transepithelial electrical resistance and disruption of the apical-basolateral intramembrane diffusion barrier by expression of a mutant tight junction membrane protein. J Cell Biol. 1996 Aug;134(4):1031–1049. doi: 10.1083/jcb.134.4.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bartles J. R., Feracci H. M., Stieger B., Hubbard A. L. Biogenesis of the rat hepatocyte plasma membrane in vivo: comparison of the pathways taken by apical and basolateral proteins using subcellular fractionation. J Cell Biol. 1987 Sep;105(3):1241–1251. doi: 10.1083/jcb.105.3.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bonilha V. L., Marmorstein A. D., Cohen-Gould L., Rodriguez-Boulan E. Apical sorting of influenza hemagglutinin by transcytosis in retinal pigment epithelium. J Cell Sci. 1997 Aug;110(Pt 15):1717–1727. doi: 10.1242/jcs.110.15.1717. [DOI] [PubMed] [Google Scholar]
  4. Brewer C. B., Roth M. G. A single amino acid change in the cytoplasmic domain alters the polarized delivery of influenza virus hemagglutinin. J Cell Biol. 1991 Aug;114(3):413–421. doi: 10.1083/jcb.114.3.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brändli A. W., Parton R. G., Simons K. Transcytosis in MDCK cells: identification of glycoproteins transported bidirectionally between both plasma membrane domains. J Cell Biol. 1990 Dec;111(6 Pt 2):2909–2921. doi: 10.1083/jcb.111.6.2909. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chen Y., Merzdorf C., Paul D. L., Goodenough D. A. COOH terminus of occludin is required for tight junction barrier function in early Xenopus embryos. J Cell Biol. 1997 Aug 25;138(4):891–899. doi: 10.1083/jcb.138.4.891. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cole N. B., Ellenberg J., Song J., DiEuliis D., Lippincott-Schwartz J. Retrograde transport of Golgi-localized proteins to the ER. J Cell Biol. 1998 Jan 12;140(1):1–15. doi: 10.1083/jcb.140.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Daro E., van der Sluijs P., Galli T., Mellman I. Rab4 and cellubrevin define different early endosome populations on the pathway of transferrin receptor recycling. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9559–9564. doi: 10.1073/pnas.93.18.9559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Evan G. I., Lewis G. K., Ramsay G., Bishop J. M. Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. 1985 Dec;5(12):3610–3616. doi: 10.1128/mcb.5.12.3610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fiedler K., Simons K. Characterization of VIP36, an animal lectin homologous to leguminous lectins. J Cell Sci. 1996 Jan;109(Pt 1):271–276. doi: 10.1242/jcs.109.1.271. [DOI] [PubMed] [Google Scholar]
  11. Furuse M., Hirase T., Itoh M., Nagafuchi A., Yonemura S., Tsukita S., Tsukita S. Occludin: a novel integral membrane protein localizing at tight junctions. J Cell Biol. 1993 Dec;123(6 Pt 2):1777–1788. doi: 10.1083/jcb.123.6.1777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Futter C. E., Connolly C. N., Cutler D. F., Hopkins C. R. Newly synthesized transferrin receptors can be detected in the endosome before they appear on the cell surface. J Biol Chem. 1995 May 5;270(18):10999–11003. doi: 10.1074/jbc.270.18.10999. [DOI] [PubMed] [Google Scholar]
  13. Garcia M., Mirre C., Quaroni A., Reggio H., Le Bivic A. GPI-anchored proteins associate to form microdomains during their intracellular transport in Caco-2 cells. J Cell Sci. 1993 Apr;104(Pt 4):1281–1290. doi: 10.1242/jcs.104.4.1281. [DOI] [PubMed] [Google Scholar]
  14. Green S. A., Plutner H., Mellman I. Biosynthesis and intracellular transport of the mouse macrophage Fc receptor. J Biol Chem. 1985 Aug 15;260(17):9867–9874. [PubMed] [Google Scholar]
  15. Guan J. L., Machamer C. E., Rose J. K. Glycosylation allows cell-surface transport of an anchored secretory protein. Cell. 1985 Sep;42(2):489–496. doi: 10.1016/0092-8674(85)90106-0. [DOI] [PubMed] [Google Scholar]
  16. Itin C., Roche A. C., Monsigny M., Hauri H. P. ERGIC-53 is a functional mannose-selective and calcium-dependent human homologue of leguminous lectins. Mol Biol Cell. 1996 Mar;7(3):483–493. doi: 10.1091/mbc.7.3.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Itin C., Schindler R., Hauri H. P. Targeting of protein ERGIC-53 to the ER/ERGIC/cis-Golgi recycling pathway. J Cell Biol. 1995 Oct;131(1):57–67. doi: 10.1083/jcb.131.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kappeler F., Itin C., Schindler R., Hauri H. P. A dual role for COOH-terminal lysine residues in pre-Golgi retention and endocytosis of ERGIC-53. J Biol Chem. 1994 Mar 4;269(9):6279–6281. [PubMed] [Google Scholar]
  19. Kappeler F., Klopfenstein D. R., Foguet M., Paccaud J. P., Hauri H. P. The recycling of ERGIC-53 in the early secretory pathway. ERGIC-53 carries a cytosolic endoplasmic reticulum-exit determinant interacting with COPII. J Biol Chem. 1997 Dec 12;272(50):31801–31808. doi: 10.1074/jbc.272.50.31801. [DOI] [PubMed] [Google Scholar]
  20. Keller P., Simons K. Post-Golgi biosynthetic trafficking. J Cell Sci. 1997 Dec;110(Pt 24):3001–3009. doi: 10.1242/jcs.110.24.3001. [DOI] [PubMed] [Google Scholar]
  21. Kundu A., Avalos R. T., Sanderson C. M., Nayak D. P. Transmembrane domain of influenza virus neuraminidase, a type II protein, possesses an apical sorting signal in polarized MDCK cells. J Virol. 1996 Sep;70(9):6508–6515. doi: 10.1128/jvi.70.9.6508-6515.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Le Bivic A., Quaroni A., Nichols B., Rodriguez-Boulan E. Biogenetic pathways of plasma membrane proteins in Caco-2, a human intestinal epithelial cell line. J Cell Biol. 1990 Oct;111(4):1351–1361. doi: 10.1083/jcb.111.4.1351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Leitinger B., Hille-Rehfeld A., Spiess M. Biosynthetic transport of the asialoglycoprotein receptor H1 to the cell surface occurs via endosomes. Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10109–10113. doi: 10.1073/pnas.92.22.10109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Linstedt A. D., Hauri H. P. Giantin, a novel conserved Golgi membrane protein containing a cytoplasmic domain of at least 350 kDa. Mol Biol Cell. 1993 Jul;4(7):679–693. doi: 10.1091/mbc.4.7.679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Low S. H., Tang B. L., Wong S. H., Hong W. Selective inhibition of protein targeting to the apical domain of MDCK cells by brefeldin A. J Cell Biol. 1992 Jul;118(1):51–62. doi: 10.1083/jcb.118.1.51. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marzolo M. P., Bull P., González A. Apical sorting of hepatitis B surface antigen (HBsAg) is independent of N-glycosylation and glycosylphosphatidylinositol-anchored protein segregation. Proc Natl Acad Sci U S A. 1997 Mar 4;94(5):1834–1839. doi: 10.1073/pnas.94.5.1834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Matter K., Balda M. S. Biogenesis of tight junctions: the C-terminal domain of occludin mediates basolateral targeting. J Cell Sci. 1998 Feb;111(Pt 4):511–519. doi: 10.1242/jcs.111.4.511. [DOI] [PubMed] [Google Scholar]
  28. Matter K., Brauchbar M., Bucher K., Hauri H. P. Sorting of endogenous plasma membrane proteins occurs from two sites in cultured human intestinal epithelial cells (Caco-2). Cell. 1990 Feb 9;60(3):429–437. doi: 10.1016/0092-8674(90)90594-5. [DOI] [PubMed] [Google Scholar]
  29. Matter K., Hunziker W., Mellman I. Basolateral sorting of LDL receptor in MDCK cells: the cytoplasmic domain contains two tyrosine-dependent targeting determinants. Cell. 1992 Nov 27;71(5):741–753. doi: 10.1016/0092-8674(92)90551-m. [DOI] [PubMed] [Google Scholar]
  30. Matter K., Mellman I. Mechanisms of cell polarity: sorting and transport in epithelial cells. Curr Opin Cell Biol. 1994 Aug;6(4):545–554. doi: 10.1016/0955-0674(94)90075-2. [DOI] [PubMed] [Google Scholar]
  31. Matter K., Whitney J. A., Yamamoto E. M., Mellman I. Common signals control low density lipoprotein receptor sorting in endosomes and the Golgi complex of MDCK cells. Cell. 1993 Sep 24;74(6):1053–1064. doi: 10.1016/0092-8674(93)90727-8. [DOI] [PubMed] [Google Scholar]
  32. Mellman I. Endocytosis and molecular sorting. Annu Rev Cell Dev Biol. 1996;12:575–625. doi: 10.1146/annurev.cellbio.12.1.575. [DOI] [PubMed] [Google Scholar]
  33. Miettinen H. M., Matter K., Hunziker W., Rose J. K., Mellman I. Fc receptor endocytosis is controlled by a cytoplasmic domain determinant that actively prevents coated pit localization. J Cell Biol. 1992 Feb;116(4):875–888. doi: 10.1083/jcb.116.4.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mirre C., Monlauzeur L., Garcia M., Delgrossi M. H., Le Bivic A. Detergent-resistant membrane microdomains from Caco-2 cells do not contain caveolin. Am J Physiol. 1996 Sep;271(3 Pt 1):C887–C894. doi: 10.1152/ajpcell.1996.271.3.C887. [DOI] [PubMed] [Google Scholar]
  35. Müsch A., Xu H., Shields D., Rodriguez-Boulan E. Transport of vesicular stomatitis virus G protein to the cell surface is signal mediated in polarized and nonpolarized cells. J Cell Biol. 1996 May;133(3):543–558. doi: 10.1083/jcb.133.3.543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Nishimura N., Balch W. E. A di-acidic signal required for selective export from the endoplasmic reticulum. Science. 1997 Jul 25;277(5325):556–558. doi: 10.1126/science.277.5325.556. [DOI] [PubMed] [Google Scholar]
  37. Odorizzi G., Trowbridge I. S. Structural requirements for basolateral sorting of the human transferrin receptor in the biosynthetic and endocytic pathways of Madin-Darby canine kidney cells. J Cell Biol. 1997 Jun 16;137(6):1255–1264. doi: 10.1083/jcb.137.6.1255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Rodriguez-Boulan E., Powell S. K. Polarity of epithelial and neuronal cells. Annu Rev Cell Biol. 1992;8:395–427. doi: 10.1146/annurev.cb.08.110192.002143. [DOI] [PubMed] [Google Scholar]
  39. Sakakibara A., Furuse M., Saitou M., Ando-Akatsuka Y., Tsukita S. Possible involvement of phosphorylation of occludin in tight junction formation. J Cell Biol. 1997 Jun 16;137(6):1393–1401. doi: 10.1083/jcb.137.6.1393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Scheiffele P., Peränen J., Simons K. N-glycans as apical sorting signals in epithelial cells. Nature. 1995 Nov 2;378(6552):96–98. doi: 10.1038/378096a0. [DOI] [PubMed] [Google Scholar]
  41. Scheiffele P., Roth M. G., Simons K. Interaction of influenza virus haemagglutinin with sphingolipid-cholesterol membrane domains via its transmembrane domain. EMBO J. 1997 Sep 15;16(18):5501–5508. doi: 10.1093/emboj/16.18.5501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schekman R., Orci L. Coat proteins and vesicle budding. Science. 1996 Mar 15;271(5255):1526–1533. doi: 10.1126/science.271.5255.1526. [DOI] [PubMed] [Google Scholar]
  43. Schweizer A., Fransen J. A., Bächi T., Ginsel L., Hauri H. P. Identification, by a monoclonal antibody, of a 53-kD protein associated with a tubulo-vesicular compartment at the cis-side of the Golgi apparatus. J Cell Biol. 1988 Nov;107(5):1643–1653. doi: 10.1083/jcb.107.5.1643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Simons K., Ikonen E. Functional rafts in cell membranes. Nature. 1997 Jun 5;387(6633):569–572. doi: 10.1038/42408. [DOI] [PubMed] [Google Scholar]
  45. Sivasubramanian N., Nayak D. P. Mutational analysis of the signal-anchor domain of influenza virus neuraminidase. Proc Natl Acad Sci U S A. 1987 Jan;84(1):1–5. doi: 10.1073/pnas.84.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Thomas D. C., Brewer C. B., Roth M. G. Vesicular stomatitis virus glycoprotein contains a dominant cytoplasmic basolateral sorting signal critically dependent upon a tyrosine. J Biol Chem. 1993 Feb 15;268(5):3313–3320. [PubMed] [Google Scholar]
  47. Wahlberg J. M., Geffen I., Reymond F., Simmen T., Spiess M. trans-Golgi retention of a plasma membrane protein: mutations in the cytoplasmic domain of the asialoglycoprotein receptor subunit H1 result in trans-Golgi retention. J Cell Biol. 1995 Jul;130(2):285–297. doi: 10.1083/jcb.130.2.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Weimbs T., Low S. H., Chapin S. J., Mostov K. E. Apical targeting in polarized epithelial cells: There's more afloat than rafts. Trends Cell Biol. 1997 Oct;7(10):393–399. doi: 10.1016/S0962-8924(97)01130-6. [DOI] [PubMed] [Google Scholar]
  49. Wessel D., Flügge U. I. A method for the quantitative recovery of protein in dilute solution in the presence of detergents and lipids. Anal Biochem. 1984 Apr;138(1):141–143. doi: 10.1016/0003-2697(84)90782-6. [DOI] [PubMed] [Google Scholar]
  50. Yeaman C., Le Gall A. H., Baldwin A. N., Monlauzeur L., Le Bivic A., Rodriguez-Boulan E. The O-glycosylated stalk domain is required for apical sorting of neurotrophin receptors in polarized MDCK cells. J Cell Biol. 1997 Nov 17;139(4):929–940. doi: 10.1083/jcb.139.4.929. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Yoshimori T., Keller P., Roth M. G., Simons K. Different biosynthetic transport routes to the plasma membrane in BHK and CHO cells. J Cell Biol. 1996 Apr;133(2):247–256. doi: 10.1083/jcb.133.2.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zurzolo C., van't Hof W., van Meer G., Rodriguez-Boulan E. VIP21/caveolin, glycosphingolipid clusters and the sorting of glycosylphosphatidylinositol-anchored proteins in epithelial cells. EMBO J. 1994 Jan 1;13(1):42–53. doi: 10.1002/j.1460-2075.1994.tb06233.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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