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J Cell Biol. 1996 May 1; 133(3): 543–558.
PMCID: PMC2120809

Transport of vesicular stomatitis virus G protein to the cell surface is signal mediated in polarized and nonpolarized cells


Current model propose that in nonpolarized cells, transport of plasma membrane proteins to the surface occurs by default. In contrast, compelling evidence indicates that in polarized epithelial cells, plasma membrane proteins are sorted in the TGN into at least two vectorial routes to apical and basolateral surface domains. Since both apical and basolateral proteins are also normally expressed by both polarized and nonpolarized cells, we explored here whether recently described basolateral sorting signals in the cytoplasmic domain of basolateral proteins are recognized and used for post TGN transport by nonpolarized cells. To this end, we compared the inhibitory effect of basolateral signal peptides on the cytosol-stimulated release of two basolateral and one apical marker in semi-intact fibroblasts (3T3), pituitary (GH3), and epithelial (MDCK) cells. A basolateral signal peptide (VSVGp) corresponding to the 29-amino acid cytoplasmic tail of vesicular stomatitis virus G protein (VSVG) inhibited with identical potency the vesicular release of VSVG from the TGN of all three cell lines. On the other hand, the VSVG peptide did not inhibit the vesicular release of HA in MDCK cells not of two polypeptide hormones (growth hormone and prolactin) in GH3 cells, whereas in 3T3 cells (influenza) hemagglutinin was inhibited, albeit with a 3x lower potency than VSVG. The results support the existence of a basolateral-like, signal-mediated constitutive pathway from TGN to plasma membrane in all three cell types, and suggest that an apical-like pathway may be present in fibroblast. The data support cargo protein involvement, not bulk flow, in the formation of post-TGN vesicles and predict the involvement of distinct cytosolic factors in the assembly of apical and basolateral transport vesicles.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Austin CD, Shields D. Prosomatostatin processing in permeabilized cells. Calcium is required for prohormone cleavage but not formation of nascent secretory vesicles. J Biol Chem. 1996 Jan 12;271(2):1194–1199. [PubMed]
  • Apodaca G, Aroeti B, Tang K, Mostov KE. Brefeldin-A inhibits the delivery of the polymeric immunoglobulin receptor to the basolateral surface of MDCK cells. J Biol Chem. 1993 Sep 25;268(27):20380–20385. [PubMed]
  • Aroeti B, Mostov KE. Polarized sorting of the polymeric immunoglobulin receptor in the exocytotic and endocytotic pathways is controlled by the same amino acids. EMBO J. 1994 May 15;13(10):2297–2304. [PMC free article] [PubMed]
  • Aroeti B, Kosen PA, Kuntz ID, Cohen FE, Mostov KE. Mutational and secondary structural analysis of the basolateral sorting signal of the polymeric immunoglobulin receptor. J Cell Biol. 1993 Dec;123(5):1149–1160. [PMC free article] [PubMed]
  • Arvan P, Castle D. Protein sorting and secretion granule formation in regulated secretory cells. Trends Cell Biol. 1992 Nov;2(11):327–331. [PubMed]
  • Bauerfeind R, Huttner WB. Biogenesis of constitutive secretory vesicles, secretory granules and synaptic vesicles. Curr Opin Cell Biol. 1993 Aug;5(4):628–635. [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]
  • Bomsel M, Mostov KE. Possible role of both the alpha and beta gamma subunits of the heterotrimeric G protein, Gs, in transcytosis of the polymeric immunoglobulin receptor. J Biol Chem. 1993 Dec 5;268(34):25824–25835. [PubMed]
  • Brewer CB, Roth MG. 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. [PMC free article] [PubMed]
  • Casanova JE, Apodaca G, Mostov KE. An autonomous signal for basolateral sorting in the cytoplasmic domain of the polymeric immunoglobulin receptor. Cell. 1991 Jul 12;66(1):65–75. [PubMed]
  • Chanat E, Huttner WB. Milieu-induced, selective aggregation of regulated secretory proteins in the trans-Golgi network. J Cell Biol. 1991 Dec;115(6):1505–1519. [PMC free article] [PubMed]
  • Chege NW, Pfeffer SR. Compartmentation of the Golgi complex: brefeldin-A distinguishes trans-Golgi cisternae from the trans-Golgi network. J Cell Biol. 1990 Sep;111(3):893–899. [PMC free article] [PubMed]
  • Chung KN, Walter P, Aponte GW, Moore HP. Molecular sorting in the secretory pathway. Science. 1989 Jan 13;243(4888):192–197. [PubMed]
  • Etchison JR, Robertson JS, Summers DF. Partial structural analysis of the oligosaccharide moieties of the vesicular stomatitis virus glycoprotein by sequential chemical and enzymatic degradation. Virology. 1977 May 15;78(2):375–392. [PubMed]
  • Gonzalez A, Rizzolo L, Rindler M, Adesnik M, Sabatini DD, Gottlieb T. Nonpolarized secretion of truncated forms of the influenza hemagglutinin and the vesicular stomatitus virus G protein from MDCK cells. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3738–3742. [PMC free article] [PubMed]
  • Green R, Shields D. Somatostatin discriminates between the intracellular pathways of secretory and membrane proteins. J Cell Biol. 1984 Jul;99(1 Pt 1):97–104. [PMC free article] [PubMed]
  • Griffiths G, Simons K. The trans Golgi network: sorting at the exit site of the Golgi complex. Science. 1986 Oct 24;234(4775):438–443. [PubMed]
  • Griffiths G, Hoflack B, Simons K, Mellman I, Kornfeld S. The mannose 6-phosphate receptor and the biogenesis of lysosomes. Cell. 1988 Feb 12;52(3):329–341. [PubMed]
  • Humphrey JS, Peters PJ, Yuan LC, Bonifacino JS. Localization of TGN38 to the trans-Golgi network: involvement of a cytoplasmic tyrosine-containing sequence. J Cell Biol. 1993 Mar;120(5):1123–1135. [PMC free article] [PubMed]
  • Hunziker W, Harter C, Matter K, Mellman I. Basolateral sorting in MDCK cells requires a distinct cytoplasmic domain determinant. Cell. 1991 Sep 6;66(5):907–920. [PubMed]
  • Kornfeld R, Kornfeld S. Assembly of asparagine-linked oligosaccharides. Annu Rev Biochem. 1985;54:631–664. [PubMed]
  • Kreis TE. Microinjected antibodies against the cytoplasmic domain of vesicular stomatitis virus glycoprotein block its transport to the cell surface. EMBO J. 1986 May;5(5):931–941. [PMC free article] [PubMed]
  • Kuliawat R, Arvan P. Distinct molecular mechanisms for protein sorting within immature secretory granules of pancreatic beta-cells. J Cell Biol. 1994 Jul;126(1):77–86. [PMC free article] [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]
  • Le Bivic A, Sambuy Y, Patzak A, Patil N, Chao M, Rodriguez-Boulan E. An internal deletion in the cytoplasmic tail reverses the apical localization of human NGF receptor in transfected MDCK cells. J Cell Biol. 1991 Nov;115(3):607–618. [PMC free article] [PubMed]
  • Lisanti MP, Rodriguez-Boulan E. Glycophospholipid membrane anchoring provides clues to the mechanism of protein sorting in polarized epithelial cells. Trends Biochem Sci. 1990 Mar;15(3):113–118. [PubMed]
  • Ludwig T, Le Borgne R, Hoflack B. Roles for mannose-6-phosphate receptors in lysosomal enzyme sorting, IGF-II binding and clathrin-coat assembly. Trends Cell Biol. 1995 May;5(5):202–206. [PubMed]
  • Malhotra V, Serafini T, Orci L, Shepherd JC, Rothman JE. Purification of a novel class of coated vesicles mediating biosynthetic protein transport through the Golgi stack. Cell. 1989 Jul 28;58(2):329–336. [PubMed]
  • Matlin KS, Simons K. Sorting of an apical plasma membrane glycoprotein occurs before it reaches the cell surface in cultured epithelial cells. J Cell Biol. 1984 Dec;99(6):2131–2139. [PMC free article] [PubMed]
  • Matter K, Mellman I. Mechanisms of cell polarity: sorting and transport in epithelial cells. Curr Opin Cell Biol. 1994 Aug;6(4):545–554. [PubMed]
  • 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. [PubMed]
  • Melançon P, Franzusoff A, Howell KE. Vesicle budding: insights from cell-free assays. Trends Cell Biol. 1991 Dec;1(6):165–171. [PubMed]
  • Moore HP, Kelly RB. Secretory protein targeting in a pituitary cell line: differential transport of foreign secretory proteins to distinct secretory pathways. J Cell Biol. 1985 Nov;101(5 Pt 1):1773–1781. [PMC free article] [PubMed]
  • Moremen KW, Touster O, Robbins PW. Novel purification of the catalytic domain of Golgi alpha-mannosidase II. Characterization and comparison with the intact enzyme. J Biol Chem. 1991 Sep 5;266(25):16876–16885. [PubMed]
  • Mostov KE, Breitfeld P, Harris JM. An anchor-minus form of the polymeric immunoglobulin receptor is secreted predominantly apically in Madin-Darby canine kidney cells. J Cell Biol. 1987 Nov;105(5):2031–2036. [PMC free article] [PubMed]
  • Nabi IR, Le Bivic A, Fambrough D, Rodriguez-Boulan E. An endogenous MDCK lysosomal membrane glycoprotein is targeted basolaterally before delivery to lysosomes. J Cell Biol. 1991 Dec;115(6):1573–1584. [PMC free article] [PubMed]
  • Orci L, Glick BS, Rothman JE. A new type of coated vesicular carrier that appears not to contain clathrin: its possible role in protein transport within the Golgi stack. Cell. 1986 Jul 18;46(2):171–184. [PubMed]
  • Orci L, Ravazzola M, Amherdt M, Perrelet A, Powell SK, Quinn DL, Moore HP. The trans-most cisternae of the Golgi complex: a compartment for sorting of secretory and plasma membrane proteins. Cell. 1987 Dec 24;51(6):1039–1051. [PubMed]
  • Orci L, Ravazzola M, Storch MJ, Anderson RG, Vassalli JD, Perrelet A. Proteolytic maturation of insulin is a post-Golgi event which occurs in acidifying clathrin-coated secretory vesicles. Cell. 1987 Jun 19;49(6):865–868. [PubMed]
  • Pimplikar SW, Simons K. Regulation of apical transport in epithelial cells by a Gs class of heterotrimeric G protein. Nature. 1993 Apr 1;362(6419):456–458. [PubMed]
  • Pimplikar SW, Ikonen E, Simons K. Basolateral protein transport in streptolysin O-permeabilized MDCK cells. J Cell Biol. 1994 Jun;125(5):1025–1035. [PMC free article] [PubMed]
  • Rajasekaran AK, Humphrey JS, Wagner M, Miesenböck G, Le Bivic A, Bonifacino JS, Rodriguez-Boulan E. TGN38 recycles basolaterally in polarized Madin-Darby canine kidney cells. Mol Biol Cell. 1994 Oct;5(10):1093–1103. [PMC free article] [PubMed]
  • Rindler MJ, Ivanov IE, Plesken H, Rodriguez-Boulan E, Sabatini DD. Viral glycoproteins destined for apical or basolateral plasma membrane domains traverse the same Golgi apparatus during their intracellular transport in doubly infected Madin-Darby canine kidney cells. J Cell Biol. 1984 Apr;98(4):1304–1319. [PMC free article] [PubMed]
  • Rodriguez-Boulan E, Powell SK. Polarity of epithelial and neuronal cells. Annu Rev Cell Biol. 1992;8:395–427. [PubMed]
  • Rodriguez Boulan E, Sabatini DD. Asymmetric budding of viruses in epithelial monlayers: a model system for study of epithelial polarity. Proc Natl Acad Sci U S A. 1978 Oct;75(10):5071–5075. [PMC free article] [PubMed]
  • Rose JK, Bergmann JE. Altered cytoplasmic domains affect intracellular transport of the vesicular stomatitis virus glycoprotein. Cell. 1983 Sep;34(2):513–524. [PubMed]
  • Roth J, Taatjes DJ, Lucocq JM, Weinstein J, Paulson JC. Demonstration of an extensive trans-tubular network continuous with the Golgi apparatus stack that may function in glycosylation. Cell. 1985 Nov;43(1):287–295. [PubMed]
  • Roth MG, Doyle C, Sambrook J, Gething MJ. Heterologous transmembrane and cytoplasmic domains direct functional chimeric influenza virus hemagglutinins into the endocytic pathway. J Cell Biol. 1986 Apr;102(4):1271–1283. [PMC free article] [PubMed]
  • Rubin LA, Kurman CC, Biddison WE, Goldman ND, Nelson DL. A monoclonal antibody 7G7/B6, binds to an epitope on the human interleukin-2 (IL-2) receptor that is distinct from that recognized by IL-2 or anti-Tac. Hybridoma. 1985 Summer;4(2):91–102. [PubMed]
  • Simons K, Wandinger-Ness A. Polarized sorting in epithelia. Cell. 1990 Jul 27;62(2):207–210. [PubMed]
  • Stoller TJ, Shields D. Retrovirus-mediated expression of preprosomatostatin: posttranslational processing, intracellular storage, and secretion in GH3 pituitary cells. J Cell Biol. 1988 Dec;107(6 Pt 1):2087–2095. [PMC free article] [PubMed]
  • Stoller TJ, Shields D. The propeptide of preprosomatostatin mediates intracellular transport and secretion of alpha-globin from mammalian cells. J Cell Biol. 1989 May;108(5):1647–1655. [PMC free article] [PubMed]
  • Tarentino AL, Trimble RB, Maley F. endo-beta-N-Acetylglucosaminidase from Streptomyces plicatus. Methods Enzymol. 1978;50:574–580. [PubMed]
  • Thomas DC, Roth MG. The basolateral targeting signal in the cytoplasmic domain of glycoprotein G from vesicular stomatitis virus resembles a variety of intracellular targeting motifs related by primary sequence but having diverse targeting activities. J Biol Chem. 1994 Jun 3;269(22):15732–15739. [PubMed]
  • Thomas DC, Brewer CB, Roth MG. 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]
  • Tooze J, Tooze SA. Clathrin-coated vesicular transport of secretory proteins during the formation of ACTH-containing secretory granules in AtT20 cells. J Cell Biol. 1986 Sep;103(3):839–850. [PMC free article] [PubMed]
  • Tooze SA, Huttner WB. Cell-free protein sorting to the regulated and constitutive secretory pathways. Cell. 1990 Mar 9;60(5):837–847. [PubMed]
  • van Meer G, Simons K. Lipid polarity and sorting in epithelial cells. J Cell Biochem. 1988 Jan;36(1):51–58. [PubMed]
  • Wandinger-Ness A, Bennett MK, Antony C, Simons K. Distinct transport vesicles mediate the delivery of plasma membrane proteins to the apical and basolateral domains of MDCK cells. J Cell Biol. 1990 Sep;111(3):987–1000. [PMC free article] [PubMed]
  • Wieland FT, Gleason ML, Serafini TA, Rothman JE. The rate of bulk flow from the endoplasmic reticulum to the cell surface. Cell. 1987 Jul 17;50(2):289–300. [PubMed]
  • Xu H, Shields D. Prohormone processing in the trans-Golgi network: endoproteolytic cleavage of prosomatostatin and formation of nascent secretory vesicles in permeabilized cells. J Cell Biol. 1993 Sep;122(6):1169–1184. [PMC free article] [PubMed]

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