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Microtubule-acting drugs lead to the nonpolarized delivery of the influenza hemagglutinin to the cell surface of polarized Madin-Darby canine kidney cells.

The Journal of Cell Biology, 01 Feb 1987, 104(2):231-241
https://doi.org/10.1083/jcb.104.2.231 PMID: 2879845 PMCID: PMC2114410

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Abstract 

The synchronized directed transfer of the envelope glycoproteins of the influenza and vesicular stomatitis viruses from the Golgi apparatus to the apical and basolateral surfaces, respectively, of polarized Madin-Darby canine kidney (MDCK) cells can be achieved using temperature-sensitive mutant viruses and appropriate temperature shift protocols (Rindler, M. J., I. E. Ivanov, H. Plesken, and D. D. Sabatini, 1985, J. Cell Biol., 100:136-151). The microtubule-depolymerizing agents colchicine and nocodazole, as well as the microtubule assembly-promoting drug taxol, were found to interfere with the normal polarized delivery and exclusive segregation of hemagglutinin (HA) to the apical surface but not with the delivery and initial accumulation of G on the basolateral surface. Immunofluorescence analysis of permeabilized monolayers of influenza-infected MDCK cells treated with the microtubule-acting drugs demonstrated the presence of substantial amounts of HA protein on both the apical and basolateral surfaces. Moreover, in cells infected with the wild-type influenza virus, particles budded from both surfaces. Viral counts in electron micrographs showed that approximately 40% of the released viral particles accumulated in the intercellular spaces or were trapped between the cell and monolayer and the collagen support as compared to less than 1% on the basolateral surface of untreated infected cells. The effect of the microtubule inhibitors was not a result of a rapid redistribution of glycoprotein molecules initially delivered to the apical surface since a redistribution was not observed when the inhibitors were added to the cells after the HA was permitted to reach the apical surface at the permissive temperature and the synthesis of new HA was inhibited with cycloheximide. The altered segregation of the HA protein that occurs may result from the dispersal of the Golgi apparatus induced by the inhibitors or from the disruption of putative microtubules containing tracks that could direct vesicles from the trans Golgi apparatus to the cell surface. Since the vesicular stomatitis virus G protein is basolaterally segregated even when the Golgi elements are dispersed and hypothetical tracks disrupted, it appears that the two viral envelope glycoproteins are segregated by fundamentally different mechanisms and that the apical surface may be incapable of accepting vesicles carrying the G protein.

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J Cell Biol. 1987 Feb 1; 104(2): 231–241.
PMCID: PMC2114410
PMID: 2879845

Microtubule-acting drugs lead to the nonpolarized delivery of the influenza hemagglutinin to the cell surface of polarized Madin-Darby canine kidney cells

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Abstract

The synchronized directed transfer of the envelope glycoproteins of the influenza and vesicular stomatitis viruses from the Golgi apparatus to the apical and basolateral surfaces, respectively, of polarized Madin- Darby canine kidney (MDCK) cells can be achieved using temperature- sensitive mutant viruses and appropriate temperature shift protocols (Rindler, M. J., I. E. Ivanov, H. Plesken, and D. D. Sabatini, 1985, J. Cell Biol., 100:136-151). The microtubule-depolymerizing agents colchicine and nocodazole, as well as the microtubule assembly- promoting drug taxol, were found to interfere with the normal polarized delivery and exclusive segregation of hemagglutinin (HA) to the apical surface but not with the delivery and initial accumulation of G on the basolateral surface. Immunofluorescence analysis of permeabilized monolayers of influenza-infected MDCK cells treated with the microtubule-acting drugs demonstrated the presence of substantial amounts of HA protein on both the apical and basolateral surfaces. Moreover, in cells infected with the wild-type influenza virus, particles budded from both surfaces. Viral counts in electron micrographs showed that approximately 40% of the released viral particles accumulated in the intercellular spaces or were trapped between the cell and monolayer and the collagen support as compared to less than 1% on the basolateral surface of untreated infected cells. The effect of the microtubule inhibitors was not a result of a rapid redistribution of glycoprotein molecules initially delivered to the apical surface since a redistribution was not observed when the inhibitors were added to the cells after the HA was permitted to reach the apical surface at the permissive temperature and the synthesis of new HA was inhibited with cycloheximide. The altered segregation of the HA protein that occurs may result from the dispersal of the Golgi apparatus induced by the inhibitors or from the disruption of putative microtubules containing tracks that could direct vesicles from the trans Golgi apparatus to the cell surface. Since the vesicular stomatitis virus G protein is basolaterally segregated even when the Golgi elements are dispersed and hypothetical tracks disrupted, it appears that the two viral envelope glycoproteins are segregated by fundamentally different mechanisms and that the apical surface may be incapable of accepting vesicles carrying the G protein.

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Selected References

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  • Bergmann JE, Kupfer A, Singer SJ. Membrane insertion at the leading edge of motile fibroblasts. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1367–1371. [Europe PMC free article] [Abstract] [Google Scholar]
  • Blok J, Ginsel LA, Mulder-Stapel AA, Onderwater JJ, Daems WT. The effect of colchicine on the intracellular transport of 3H-fucose-labelled glycoproteins in the absorptive cells of cultured human small-intestinal tissue. An autoradiographical and biochemical study. Cell Tissue Res. 1981;215(1):1–12. [Abstract] [Google Scholar]
  • Brown JC, Salomonsky NL. Site-specific maturation of enveloped viruses in L cells treated with cytochalasin B. J Cell Biol. 1985 Feb;100(2):357–363. [Europe PMC free article] [Abstract] [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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Danielsen EM, Cowell GM, Poulsen SS. Biosynthesis of intestinal microvillar proteins. Role of the Golgi complex and microtubules. Biochem J. 1983 Oct 15;216(1):37–42. [Europe PMC free article] [Abstract] [Google Scholar]
  • De Brabander M, Geuens G, Nuydens R, Willebrords R, De Mey J. Taxol induces the assembly of free microtubules in living cells and blocks the organizing capacity of the centrosomes and kinetochores. Proc Natl Acad Sci U S A. 1981 Sep;78(9):5608–5612. [Europe PMC free article] [Abstract] [Google Scholar]
  • De Brabander MJ, Van de Veire RM, Aerts FE, Borgers M, Janssen PA. The effects of methyl (5-(2-thienylcarbonyl)-1H-benzimidazol-2-yl) carbamate, (R 17934; NSC 238159), a new synthetic antitumoral drug interfering with microtubules, on mammalian cells cultured in vitro. Cancer Res. 1976 Mar;36(3):905–916. [Abstract] [Google Scholar]
  • Ellinger A, Pavelka M, Gangl A. Effect of colchicine on rat small intestinal absorptive cells. II. Distribution of label after incorporation of [3H]fucose into plasma membrane glycoproteins. J Ultrastruct Res. 1983 Dec;85(3):260–271. [Abstract] [Google Scholar]
  • Griffiths G, Pfeiffer S, Simons K, Matlin K. Exit of newly synthesized membrane proteins from the trans cisterna of the Golgi complex to the plasma membrane. J Cell Biol. 1985 Sep;101(3):949–964. [Europe PMC free article] [Abstract] [Google Scholar]
  • Louvard D. Apical membrane aminopeptidase appears at site of cell-cell contact in cultured kidney epithelial cells. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4132–4136. [Europe PMC free article] [Abstract] [Google Scholar]
  • Louvard D, Reggio H, Warren G. Antibodies to the Golgi complex and the rough endoplasmic reticulum. J Cell Biol. 1982 Jan;92(1):92–107. [Europe PMC free article] [Abstract] [Google Scholar]
  • MARCUS PI. Dynamics of surface modification in myxovirus-infected cells. Cold Spring Harb Symp Quant Biol. 1962;27:351–365. [Abstract] [Google Scholar]
  • Matlin KS, 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. [Abstract] [Google Scholar]
  • Meza I, Ibarra G, Sabanero M, Martínez-Palomo A, Cereijido M. Occluding junctions and cytoskeletal components in a cultured transporting epithelium. J Cell Biol. 1980 Dec;87(3 Pt 1):746–754. [Europe PMC free article] [Abstract] [Google Scholar]
  • Murphy DB, Tilney LG. The role of microtubules in the movement of pigment granules in teleost melanophores. J Cell Biol. 1974 Jun;61(3):757–779. [Europe PMC free article] [Abstract] [Google Scholar]
  • Nagura H, Nakane PK, Brown WR. Translocation of dimeric IgA through neoplastic colon cells in vitro. J Immunol. 1979 Nov;123(5):2359–2368. [Abstract] [Google Scholar]
  • Pavelka M, Ellinger A, Gangl A. Effect of colchicine on rat small intestinal absorptive cells. I Formation of basolateral microvillus borders. J Ultrastruct Res. 1983 Dec;85(3):249–259. [Abstract] [Google Scholar]
  • Quaroni A, Kirsch K, Weiser MM. Synthesis of membrane glycoproteins in rat small-intestinal villus cells. Effect of colchicine on the redistribution of L-[1,5,6-3H]fucose-labelled membrane glycoproteins among Golgi, lateral basal and microvillus membranes. Biochem J. 1979 Jul 15;182(1):213–221. [Europe PMC free article] [Abstract] [Google Scholar]
  • Redman CM, Banerjee D, Howell K, Palade GE. Colchicine inhibition of plasma protein release from rat hepatocytes. J Cell Biol. 1975 Jul;66(1):42–59. [Europe PMC free article] [Abstract] [Google Scholar]
  • 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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rindler MJ, Ivanov IE, Plesken H, Sabatini DD. Polarized delivery of viral glycoproteins to the apical and basolateral plasma membranes of Madin-Darby canine kidney cells infected with temperature-sensitive viruses. J Cell Biol. 1985 Jan;100(1):136–151. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rodriguez Boulan E, Pendergast M. Polarized distribution of viral envelope proteins in the plasma membrane of infected epithelial cells. Cell. 1980 May;20(1):45–54. [Abstract] [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. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rodriguez-Boulan E, Paskiet KT, Salas PJ, Bard E. Intracellular transport of influenza virus hemagglutinin to the apical surface of Madin-Darby canine kidney cells. J Cell Biol. 1984 Jan;98(1):308–319. [Europe PMC free article] [Abstract] [Google Scholar]
  • Rogalski AA, Bergmann JE, Singer SJ. Effect of microtubule assembly status on the intracellular processing and surface expression of an integral protein of the plasma membrane. J Cell Biol. 1984 Sep;99(3):1101–1109. [Europe PMC free article] [Abstract] [Google Scholar]
  • Salas PJ, Misek DE, Vega-Salas DE, Gundersen D, Cereijido M, Rodriguez-Boulan E. Microtubules and actin filaments are not critically involved in the biogenesis of epithelial cell surface polarity. J Cell Biol. 1986 May;102(5):1853–1867. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schiff PB, Horwitz SB. Taxol stabilizes microtubules in mouse fibroblast cells. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1561–1565. [Europe PMC free article] [Abstract] [Google Scholar]
  • Schiff PB, Fant J, Horwitz SB. Promotion of microtubule assembly in vitro by taxol. Nature. 1979 Feb 22;277(5698):665–667. [Abstract] [Google Scholar]
  • Schnapp BJ, Vale RD, Sheetz MP, Reese TS. Single microtubules from squid axoplasm support bidirectional movement of organelles. Cell. 1985 Feb;40(2):455–462. [Abstract] [Google Scholar]
  • Ueda M, Kilbourne ED. Temperature-sensitive mutants of influenza virus: a mutation in the hemagglutinin gene. Virology. 1976 Apr;70(2):425–431. [Abstract] [Google Scholar]
  • Wehland J, Henkart M, Klausner R, Sandoval IV. Role of microtubules in the distribution of the Golgi apparatus: effect of taxol and microinjected anti-alpha-tubulin antibodies. Proc Natl Acad Sci U S A. 1983 Jul;80(14):4286–4290. [Europe PMC free article] [Abstract] [Google Scholar]
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