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. 1983 Mar 1;96(3):866–874. doi: 10.1083/jcb.96.3.866

Assembly of enveloped viruses in Madin-Darby canine kidney cells: polarized budding from single attached cells and from clusters of cells in suspension

PMCID: PMC2112422  PMID: 6300140

Abstract

In confluent monolayers of the dog kidney epithelial cell line Madin- Darby canine kidney (MDCK) assembly of RNA enveloped viruses reflects the functional polarization of the cells. Thus, influenza, Sendai, and Simian virus 5 bud from the apical (free) surface, while vesicular stomatitis virions (VSV) are assembled at basolateral plasma membrane domains (Rodriguez-Boulan, E., and D.D. Sabatini, 1978, Proc. Natl. Acad. Sci. U.S.A., 75:5071-5075). MDCK cells derived from confluent monolayers by dissociation with trypsin-EDTA and maintained as single cells in spinner medium for 12-20 h before infection, lose their characteristic structural polarity. Furthermore, when these cells were infected with influenza or VSV, virions assembled in a nonpolarized fashion over most of the cell surface. However, when dissociated MDCK cells infected in suspension were sparsely plated on collagen gels to prevent intercellular contact and the formation of junctions, the characteristic polarity of viral budding observed in confluent monolayers was again manifested; i.e., VSV budded preferentially from adherent surfaces and influenza almost exclusively from free surface regions. Similar polarization was observed in cells which became aggregated during incubation in spinner medium: influenza budded from the free surface, while VSV was produced at regions of cell-cell contact. It therefore appears that in isolated epithelial cells attachment to a substrate or to another cell is sufficient to trigger the expression of plasma membrane polarity which is manifested in the asymmetric budding of viruses.

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Selected References

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

  1. Cereijido M., Ehrenfeld J., Meza I., Martínez-Palomo A. Structural and functional membrane polarity in cultured monolayers of MDCK cells. J Membr Biol. 1980;52(2):147–159. doi: 10.1007/BF01869120. [DOI] [PubMed] [Google Scholar]
  2. Cereijido M., Robbins E. S., Dolan W. J., Rotunno C. A., Sabatini D. D. Polarized monolayers formed by epithelial cells on a permeable and translucent support. J Cell Biol. 1978 Jun;77(3):853–880. doi: 10.1083/jcb.77.3.853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dragsten P. R., Blumenthal R., Handler J. S. Membrane asymmetry in epithelia: is the tight junction a barrier to diffusion in the plasma membrane? Nature. 1981 Dec 24;294(5843):718–722. doi: 10.1038/294718a0. [DOI] [PubMed] [Google Scholar]
  4. FARQUHAR M. G., PALADE G. E. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. doi: 10.1083/jcb.17.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Goodenough D. A., Revel J. P. A fine structural analysis of intercellular junctions in the mouse liver. J Cell Biol. 1970 May;45(2):272–290. doi: 10.1083/jcb.45.2.272. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Grinnell F. Fibroblast receptor for cell-substratum adhesion: studies on the interaction of baby hamster kidney cells with latex beads coated by cold insoluble globulin (plasma fibronectin). J Cell Biol. 1980 Jul;86(1):104–112. doi: 10.1083/jcb.86.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hoi Sang U., Saier M. H., Jr, Ellisman M. H. Tight junction formation is closely linked to the polar redistribution of intramembranous particles in aggregating MDCK epithelia. Exp Cell Res. 1979 Sep;122(2):384–391. doi: 10.1016/0014-4827(79)90315-x. [DOI] [PubMed] [Google Scholar]
  8. Leighton J., Brada Z., Estes L. W., Justh G. Secretory activity and oncogenicity of a cell line (MDCK) derived from canine kidney. Science. 1969 Jan 31;163(3866):472–473. doi: 10.1126/science.163.3866.472. [DOI] [PubMed] [Google Scholar]
  9. 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. doi: 10.1073/pnas.77.7.4132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Martinez-Palomo A., Meza I., Beaty G., Cereijido M. Experimental modulation of occluding junctions in a cultured transporting epithelium. J Cell Biol. 1980 Dec;87(3 Pt 1):736–745. doi: 10.1083/jcb.87.3.736. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Mauchamp J., Margotat A., Chambard M., Charrier B., Remy L., Michel-Bechet M. Polarity of three-dimensional structures derived from isolated hog thyroid cells in primary culture. Cell Tissue Res. 1979;204(3):417–430. doi: 10.1007/BF00233653. [DOI] [PubMed] [Google Scholar]
  12. Meldolesi J., Castiglioni G., Parma R., Nassivera N., De Camilli P. Ca++-dependent disassembly and reassembly of occluding junctions in guinea pig pancreatic acinar cells. Effect of drugs. J Cell Biol. 1978 Oct;79(1):156–172. doi: 10.1083/jcb.79.1.156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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. doi: 10.1083/jcb.87.3.746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Michl J., Pieczonka M. M., Unkeless J. C., Silverstein S. C. Effects of immobilized immune complexes on Fc- and complement-receptor function in resident and thioglycollate-elicited mouse peritoneal macrophages. J Exp Med. 1979 Sep 19;150(3):607–621. doi: 10.1084/jem.150.3.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Misfeldt D. S., Hamamoto S. T., Pitelka D. R. Transepithelial transport in cell culture. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1212–1216. doi: 10.1073/pnas.73.4.1212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Nitsch L., Wollman S. H. Ultrastructure of intermediate stages in polarity reversal of thyroid epithelium in follicles in suspension culture. J Cell Biol. 1980 Sep;86(3):875–880. doi: 10.1083/jcb.86.3.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pisam M., Ripoche P. Redistribution of surface macromolecules in dissociated epithelial cells. J Cell Biol. 1976 Dec;71(3):907–920. doi: 10.1083/jcb.71.3.907. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rodriguez Boulan E., Sabatini D. D. 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. doi: 10.1073/pnas.75.10.5071. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Roth M. G., Compans R. W. Delayed appearance of pseudotypes between vesicular stomatitis virus influenza virus during mixed infection of MDCK cells. J Virol. 1981 Dec;40(3):848–860. doi: 10.1128/jvi.40.3.848-860.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Weigel P. H. Rat hepatocytes bind to synthetic galactoside surfaces via a patch of asialoglycoprotein receptors. J Cell Biol. 1980 Dec;87(3 Pt 1):855–861. doi: 10.1083/jcb.87.3.855. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ziomek C. A., Schulman S., Edidin M. Redistribution of membrane proteins in isolated mouse intestinal epithelial cells. J Cell Biol. 1980 Sep;86(3):849–857. doi: 10.1083/jcb.86.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]

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