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. 1977 Aug;74(8):3433–3437. doi: 10.1073/pnas.74.8.3433

Changes in surface properties of normal and transformed cells caused by tunicamycin, an inhibitor of protein glycosylation.

D Duksin, P Bornstein
PMCID: PMC431594  PMID: 198786

Abstract

Normal and virally transformed mouse (3T3) and human (WI-38) cells were treated with tunicamycin, an inhibitor of lipid-carrier-dependent glycosylation of proteins. Incubation of cells with tunicamycin (1 microgram/ml) caused detachment and death of simian virus 40- and polyoma-transformed cells within 24 hr; these effects were not seen with nontransformed cell lines. However, the proliferation of 3T3 cells was inhibited by tunicamycin and, after a few days, a distinct change from an epithelioid to an abnormally elongated shape was observed. Both inhibition of growth and the morphological changes were reversible. A marked decrease in concanavalin A agglutinability was observed in virally transformed cells treated with tunicamycin (0.5 microgram/ml), but agglutination by wheat germ agglutinin or soybean agglutinin was unaffected. Analysis of biosynthetically labeled proteins showed that a high-molecular-weight protein, presumed to be related to fibronectin, is markedly reduced in the medium of cells cultured in the presence of tunicamycin. These results suggest that tunicamycin interferes with the insertion or function of one or more cell-surface glycoproteins. Such cell-surface changes could affect a number of cellular properties, including attachment, cell shape, and agglutinability by some lectins.

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Selected References

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

  1. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  2. Bornstein P., Von der Mark K., Wyke A. W., Ehrlich H. P., Monson J. M. Characterization of the pro- 1 chain of procollagen. J Biol Chem. 1972 May 10;247(9):2808–2813. [PubMed] [Google Scholar]
  3. Briles E. B., Li E., Kornfeld S. Isolation of wheat germ agglutinin-resistant clones of Chinese hamster ovary cells deficient in membrane sialic acid and galactose. J Biol Chem. 1977 Feb 10;252(3):1107–1116. [PubMed] [Google Scholar]
  4. Critchley D. R., Wyke J. A., Hynes R. O. Cell surface and metabolic labelling of the proteins of normal and transformed chicken cells. Biochim Biophys Acta. 1976 Jun 17;436(2):335–352. doi: 10.1016/0005-2736(76)90198-x. [DOI] [PubMed] [Google Scholar]
  5. Cuatrecasas P., Hollenberg M. D. Membrane receptors and hormone action. Adv Protein Chem. 1976;30:251–451. doi: 10.1016/s0065-3233(08)60481-7. [DOI] [PubMed] [Google Scholar]
  6. Duksin D., Bornstein P. Impaired conversion of procollagen to collagen by fibroblasts and bone treated with tunicamycin, an inhibitor of protein glycosylation. J Biol Chem. 1977 Feb 10;252(3):955–962. [PubMed] [Google Scholar]
  7. Duksin D., Katchalski E., Sachs L. Specific aggregation of SV40-transformed cells by ornithine, leucine copolymers. Proc Natl Acad Sci U S A. 1970 Sep;67(1):185–192. doi: 10.1073/pnas.67.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dulbecco R. Topoinhibition and serum requirement of transformed and untransformed cells. Nature. 1970 Aug 22;227(5260):802–806. doi: 10.1038/227802a0. [DOI] [PubMed] [Google Scholar]
  9. Edelman G. M. Surface modulation in cell recognition and cell growth. Science. 1976 Apr 16;192(4236):218–226. doi: 10.1126/science.769162. [DOI] [PubMed] [Google Scholar]
  10. Edwards J. G., Dysart J. M., Hughes R. C. Cellular adhesiveness reduced in ricin-resistant hamster fibroblasts. Nature. 1976 Nov 4;264(5581):66–68. doi: 10.1038/264066a0. [DOI] [PubMed] [Google Scholar]
  11. Hynes R. O. Cell surface proteins and malignant transformation. Biochim Biophys Acta. 1976 Apr 30;458(1):73–107. doi: 10.1016/0304-419x(76)90015-9. [DOI] [PubMed] [Google Scholar]
  12. Kruse N. J., Bornstein P. The metabolic requirements for transcellular movement and secretion of collagen. J Biol Chem. 1975 Jul 10;250(13):4841–4847. [PubMed] [Google Scholar]
  13. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  14. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  15. Lehle L., Tanner W. The specific site of tunicamycin inhibition in the formation of dolichol-bound N-acetylglucosamine derivatives. FEBS Lett. 1976 Nov 15;72(1):167–170. doi: 10.1016/0014-5793(76)80922-2. [DOI] [PubMed] [Google Scholar]
  16. Lis H., Sharon N. The biochemistry of plant lectins (phytohemagglutinins). Annu Rev Biochem. 1973;42(0):541–574. doi: 10.1146/annurev.bi.42.070173.002545. [DOI] [PubMed] [Google Scholar]
  17. Nicolson G. L. Trans-membrane control of the receptors on normal and tumor cells. II. Surface changes associated with transformation and malignancy. Biochim Biophys Acta. 1976 Apr 30;458(1):1–72. doi: 10.1016/0304-419x(76)90014-7. [DOI] [PubMed] [Google Scholar]
  18. Nicolson G. L. Transmembrane control of the receptors on normal and tumor cells. I. Cytoplasmic influence over surface components. Biochim Biophys Acta. 1976 Apr 13;457(1):57–108. doi: 10.1016/0304-4157(76)90014-9. [DOI] [PubMed] [Google Scholar]
  19. OYAMA V. I., EAGLE H. Measurement of cell growth in tissue culture with a phenol reagent (folin-ciocalteau). Proc Soc Exp Biol Med. 1956 Feb;91(2):305–307. doi: 10.3181/00379727-91-22245. [DOI] [PubMed] [Google Scholar]
  20. Pouysségur J. M., Pastan I. Mutants of Balb/c 3T3 fibroblasts defective in adhesiveness to substratum: evidence for alteration in cell surface proteins. Proc Natl Acad Sci U S A. 1976 Feb;73(2):544–548. doi: 10.1073/pnas.73.2.544. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pouysségur J., Willingham M., Pastan I. Role of cell surface carbohydrates and proteins in cell behavior: studies on the biochemical reversion of an N-acetylglucosamine-deficient fibroblast mutant. Proc Natl Acad Sci U S A. 1977 Jan;74(1):243–247. doi: 10.1073/pnas.74.1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schwarz R. T., Rohrschneider J. M., Schmidt M. F. Suppression of glycoprotein formation of Semliki Forest, influenza, and avian sarcoma virus by tunicamycin. J Virol. 1976 Sep;19(3):782–791. doi: 10.1128/jvi.19.3.782-791.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shin S. I., Freedman V. H., Risser R., Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. doi: 10.1073/pnas.72.11.4435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Struck D. K., Lennarz W. J. Evidence for the participation of saccharide-lipids in the synthesis of the oligosaccharide chain of ovalbumin. J Biol Chem. 1977 Feb 10;252(3):1007–1013. [PubMed] [Google Scholar]
  25. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  26. Takatsuki A., Tamura G. Effect of tunicamycin on the synthesis of macromolecules in cultures of chick embryo fibroblasts infected with Newcastle disease virus. J Antibiot (Tokyo) 1971 Nov;24(11):785–794. doi: 10.7164/antibiotics.24.785. [DOI] [PubMed] [Google Scholar]
  27. Tkacz J. S., 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. doi: 10.1016/s0006-291x(75)80086-6. [DOI] [PubMed] [Google Scholar]
  28. Vaheri A., Ruoslahti E. Disappearance of a major cell-type specific surface glycoprotein antigen (SF) after transformation of fibroblasts by Rous sarcoma virus. Int J Cancer. 1974 May 15;13(5):579–586. doi: 10.1002/ijc.2910130502. [DOI] [PubMed] [Google Scholar]
  29. Vlodavsky I., Sachs L. Restriction of receptor mobility and the agglutination of cells by concanavalin A. Exp Cell Res. 1975 Nov;96(1):202–214. doi: 10.1016/s0014-4827(75)80052-8. [DOI] [PubMed] [Google Scholar]
  30. Waechter C. J., Lennarz W. J. The role of polyprenol-linked sugars in glycoprotein synthesis. Annu Rev Biochem. 1976;45:95–112. doi: 10.1146/annurev.bi.45.070176.000523. [DOI] [PubMed] [Google Scholar]
  31. Warren L., Fuhrer J. P., Buck C. A. Surface glycoproteins of cells before and after transformation by oncogenic viruses. Fed Proc. 1973 Jan;32(1):80–85. [PubMed] [Google Scholar]
  32. Willingham M. C., Carchman R. A., Pastan I. H. A mutant of 3T3 cells with cyclic AMP metabolism sensitive to temperature change. Proc Natl Acad Sci U S A. 1973 Oct;70(10):2906–2910. doi: 10.1073/pnas.70.10.2906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yamada K. M., Yamada S. S., Pastan I. Cell surface protein partially restores morphology, adhesiveness, and contact inhibition of movement to transformed fibroblasts. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1217–1221. doi: 10.1073/pnas.73.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]

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