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Proc Natl Acad Sci U S A. 1979 Dec; 76(12): 6430–6434.
doi: 10.1073/pnas.76.12.6430
PMCID: PMC411878
PMID: 230510

Polarity of influenza and vesicular stomatitis virus maturation in MDCK cells: lack of a requirement for glycosylation of viral glycoproteins.

M G Roth, J P Fitzpatrick, and R W Compans
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Abstract

We have investigated whether glycosylation of membrane glycoproteins is a determinant of the site of maturation of enveloped viruses in Madin-Darby canine kidney (MDCK) cells. In MDCK cell monolayers, vesicular stomatitis virus buds exclusively from the basal or lateral plasma membranes and contains a sialylated glycoprotein, whereas influenza virus buds exclusively from the apical plasma membrane and lacks neuraminic acid. In order to study the possible relationship between glycosylation of viral glycoproteins and the budding site, infected MDCK cells were treated with tunicamycin at a concentration that completely inhibits glycosylation of viral glycoproteins and the site of virus maturation was examined by electron microscopy. When tunicamycin-treated monolayers were compared to controls, the polarity in the maturation sites of both viruses was maintained. These results indicate that glycosylation of viral glycoproteins is not required for the determination of the cellular maturation site of these enveloped viruses.

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Selected References

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  • MURPHY JS, BANG FB. Observations with the electron microscope on cells of the chick chorio-allantoic membrane infected with influenza virus. J Exp Med. 1952 Mar;95(3):259–268. [PMC free article] [PubMed] [Google Scholar]
  • 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] [Google Scholar]
  • Leighton J, Estes LW, Mansukhani S, Brada Z. A cell line derived from normal dog kidney (MDCK) exhibiting qualities of papillary adenocarcinoma and of renal tubular epithelium. Cancer. 1970 Nov;26(5):1022–1028. [PubMed] [Google Scholar]
  • Misfeldt DS, Hamamoto ST, Pitelka DR. Transepithelial transport in cell culture. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212–1216. [PMC free article] [PubMed] [Google Scholar]
  • Cereijido M, Robbins ES, Dolan WJ, Rotunno CA, Sabatini DD. Polarized monolayers formed by epithelial cells on a permeable and translucent support. J Cell Biol. 1978 Jun;77(3):853–880. [PMC free article] [PubMed] [Google Scholar]
  • Lever JE. Inducers of mammalian cell differentiation stimulate dome formation in a differentiated kidney epithelial cell line (MDCK). Proc Natl Acad Sci U S A. 1979 Mar;76(3):1323–1327. [PMC free article] [PubMed] [Google Scholar]
  • Ernst SA, Mills JW. Basolateral plasma membrane localiztion of ouabain-sensitive sodium transport sites in the secretory epithelium of the avian salt gland. J Cell Biol. 1977 Oct;75(1):74–94. [PMC free article] [PubMed] [Google Scholar]
  • Klenk HD, Choppin PW. Glycosphingolipids of plasma membranes of cultured cells and an enveloped virus (SV5) grown in these cells. Proc Natl Acad Sci U S A. 1970 May;66(1):57–64. [PMC free article] [PubMed] [Google Scholar]
  • Klenk HD, Compans RW, Choppin WP. An electron microscopic study of the presence or absence of neuraminic acid in enveloped viruses. Virology. 1970 Dec;42(4):1158–1162. [PubMed] [Google Scholar]
  • Burge BW, Huang AS. Comparison of membrane protein glycopeptides of Sindbis virus and vesicular stomatitis virus. J Virol. 1970 Aug;6(2):176–182. [PMC free article] [PubMed] [Google Scholar]
  • Tkacz JS, 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. [PubMed] [Google Scholar]
  • Lennarz WJ. Lipid linked sugars in glycoprotein synthesis. Science. 1975 Jun 6;188(4192):986–991. [PubMed] [Google Scholar]
  • Nakamura K, Compans RW. Effects of glucosamine, 2-deoxyglucose, and tunicamycin on glycosylation, sulfation, and assembly of influenza viral proteins. Virology. 1978 Feb;84(2):303–319. [PubMed] [Google Scholar]
  • Gibson R, Leavitt R, Kornfeld S, Schlesinger S. Synthesis and infectivity of vesicular stomatitis virus containing nonglycosylated G protein. Cell. 1978 Apr;13(4):671–679. [PubMed] [Google Scholar]
  • Holmes KV, Choppin PW. On the role of the response of the cell membrane in determining virus virulence. Contrasting effects of the parainfluenza virus SV5 in two cell types. J Exp Med. 1966 Sep 1;124(3):501–520. [PMC free article] [PubMed] [Google Scholar]
  • Choppin PW. Replication of influenza virus in a continuous cell line: high yield of infective virus from cells inoculated at high multiplicity. Virology. 1969 Sep;39(1):130–134. [PubMed] [Google Scholar]
  • Tobita K, Sugiura A, Enomote C, Furuyama M. Plaque assay and primary isolation of influenza A viruses in an established line of canine kidney cells (MDCK) in the presence of trypsin. Med Microbiol Immunol. 1975 Dec 30;162(1):9–14. [PubMed] [Google Scholar]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed] [Google Scholar]
  • Bonner WM, Laskey RA. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. [PubMed] [Google Scholar]
  • Holmes KV, Choppin PW. On the role of microtubules in movement and alignment of nuclei in virus-induced syncytia. J Cell Biol. 1968 Dec;39(3):526–543. [PMC free article] [PubMed] [Google Scholar]
  • Eylar EH. On the biological role of glycoproteins. J Theor Biol. 1966 Jan;10(1):89–113. [PubMed] [Google Scholar]
  • Schwarz RT, Rohrschneider JM, Schmidt MF. Suppression of glycoprotein formation of Semliki Forest, influenza, and avian sarcoma virus by tunicamycin. J Virol. 1976 Sep;19(3):782–791. [PMC free article] [PubMed] [Google Scholar]
  • Klenk HD, Wöllert W, Rott R, Scholtissek C. Association of influenza virus proteins with cytoplasmic fractions. Virology. 1974 Jan;57(1):28–41. [PubMed] [Google Scholar]
  • Gandini Attardi D, Martini G, Mattoccia E, Tocchini-Valentini GP. Effect of Xenopus laevis oocyte extract on supercoiled simian virus 40 DNA: formation of complex DNA. Proc Natl Acad Sci U S A. 1976 Feb;73(2):554–558. [PMC free article] [PubMed] [Google Scholar]
  • Duksin D, Bornstein P. Changes in surface properties of normal and transformed cells caused by tunicamycin, an inhibitor of protein glycosylation. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3433–3437. [PMC free article] [PubMed] [Google Scholar]
  • Olden K, Pratt RM, Jaworski C, Yamada KM. Evidence for role of glycoprotein carbohydrates in membrane transport: specific inhibition by tunicamycin. Proc Natl Acad Sci U S A. 1979 Feb;76(2):791–795. [PMC free article] [PubMed] [Google Scholar]
  • Klenk HD, Scholtissek C, Rott R. Inhibition of glycoprotein biosynthesis of influenza virus by D-glucosamine and 2-deoxy-D-glucose. Virology. 1972 Sep;49(3):723–734. [PubMed] [Google Scholar]
  • Gandhi SS, Stanley P, Taylor JM, White DO. Inhibition of influenza viral glycoprotein synthesis by sugars. Microbios. 1972 Jan;5(17):41–50. [PubMed] [Google Scholar]
  • Gibson R, Schlesinger S, Kornfeld S. The nonglycosylated glycoprotein of vesicular stomatitis virus is temperature-sensitive and undergoes intracellular aggregation at elevated temperatures. J Biol Chem. 1979 May 10;254(9):3600–3607. [PubMed] [Google Scholar]
  • Pouysségur J, Yamada KM. Isolation and immunological characterization of a glucose-regulated fibroblast cell surface glycoprotein and its nonglycosylated precursor. Cell. 1978 Jan;13(1):139–140. [PubMed] [Google Scholar]
  • Choppin PW, Compans RW. Phenotypic mixing of envelope proteins of the parainfluenza virus SV5 and vesicular stomatitis virus. J Virol. 1970 May;5(5):609–616. [PMC free article] [PubMed] [Google Scholar]
  • McSharry JJ, Compans RW, Choppin PW. Proteins of vesicular stomatitis virus and of phenotypically mixed vesicular stomatitis virus-simian virus 5 virions. J Virol. 1971 Nov;8(5):722–729. [PMC free article] [PubMed] [Google Scholar]
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