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. 1983 Nov;42(2):771–777. doi: 10.1128/iai.42.2.771-777.1983

Entry of Shigella dysenteriae toxin into HeLa cells.

K Eiklid, S Olsnes
PMCID: PMC264496  PMID: 6358031

Abstract

The rate of shigella toxin entry into the cytosol of HeLa S3 cells was estimated from the toxin-induced reduction in protein synthesis. Whereas high toxin concentrations strongly reduced protein synthesis within 30 min, lower concentrations required longer times. The major part of the cell-bound toxin entered only after several hours. Toxin entered cells after incubation at 25 degrees C but not at 20 degrees C, although toxin binding was the same at the two temperatures. Increasing the KCl concentration to 0.2 M protected against toxin. The toxin entry was strongly reduced when the level of ATP in the cells was reduced by incubation with metabolic inhibitors. Lysosomotrophic agents such as NH4Cl and chloroquine had little or no protective effect, but the protonophores carbonyl cyanide p-trifluoromethoxyphenylhydrazone and carbonyl cyanide m-chlorophenylhydrazone and the ionophore monensin protected cells against the toxin. Cells were also protected when the pH was reduced to 6.4. The entry of shigella toxin is discussed in relation to that of other protein toxins with intracellular sites of action.

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Selected References

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

  1. Baenziger J. U., Fiete D. Recycling of the hepatocyte asialoglycoprotein receptor does not require delivery of ligand to lysosomes. J Biol Chem. 1982 Jun 10;257(11):6007–6009. [PubMed] [Google Scholar]
  2. Brown J. E., Ussery M. A., Leppla S. H., Rothman S. W. Inhibition of protein synthesis by Shiga toxin: activation of the toxin and inhibition of peptide elongation. FEBS Lett. 1980 Aug 11;117(1):84–88. doi: 10.1016/0014-5793(80)80918-5. [DOI] [PubMed] [Google Scholar]
  3. Draper R. K., Simon M. I. The entry of diphtheria toxin into the mammalian cell cytoplasm: evidence for lysosomal involvement. J Cell Biol. 1980 Dec;87(3 Pt 1):849–854. doi: 10.1083/jcb.87.3.849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dunn W. A., Hubbard A. L., Aronson N. N., Jr Low temperature selectively inhibits fusion between pinocytic vesicles and lysosomes during heterophagy of 125I-asialofetuin by the perfused rat liver. J Biol Chem. 1980 Jun 25;255(12):5971–5978. [PubMed] [Google Scholar]
  5. Eiklid K., Olsnes S. Animal toxicity of Shigella dysenteriae cytotoxin: evidence that the neurotoxic, enterotoxic, and cytotoxic activities are due to one toxin. J Immunol. 1983 Jan;130(1):380–384. [PubMed] [Google Scholar]
  6. Eiklid K., Olsnes S. Interaction of Shigella shigae cytotoxin with receptors on sensitive and insensitive cells. J Recept Res. 1980;1(2):199–213. doi: 10.3109/10799898009044098. [DOI] [PubMed] [Google Scholar]
  7. Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  8. Marnell M. H., Stookey M., Draper R. K. Monensin blocks the transport of diphtheria toxin to the cell cytoplasm. J Cell Biol. 1982 Apr;93(1):57–62. doi: 10.1083/jcb.93.1.57. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Middlebrook J. L., Dorland R. B. Differential chemical protection of mammalian cells from the exotoxins of Corynebacterium diphtheriae and Pseudomonas aeruginosa. Infect Immun. 1977 Apr;16(1):232–239. doi: 10.1128/iai.16.1.232-239.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. O'Brien A. D., LaVeck G. D. Purification and characterization of a Shigella dysenteriae 1-like toxin produced by Escherichia coli. Infect Immun. 1983 May;40(2):675–683. doi: 10.1128/iai.40.2.675-683.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ohkuma S., Poole B. Cytoplasmic vacuolation of mouse peritoneal macrophages and the uptake into lysosomes of weakly basic substances. J Cell Biol. 1981 Sep;90(3):656–664. doi: 10.1083/jcb.90.3.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Olsnes S., Reisbig R., Eiklid K. Subunit structure of Shigella cytotoxin. J Biol Chem. 1981 Aug 25;256(16):8732–8738. [PubMed] [Google Scholar]
  13. Ray B., Wu H. C. Enhancement of cytotoxicities of ricin and Pseudomonas toxin in Chinese hamster ovary cells by nigericin. Mol Cell Biol. 1981 Jun;1(6):552–559. doi: 10.1128/mcb.1.6.552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Reisbig R., Olsnes S., Eiklid K. The cytotoxic activity of Shigella toxin. Evidence for catalytic inactivation of the 60 S ribosomal subunit. J Biol Chem. 1981 Aug 25;256(16):8739–8744. [PubMed] [Google Scholar]
  15. Sandvig K., Olsnes S. Diphtheria toxin entry into cells is facilitated by low pH. J Cell Biol. 1980 Dec;87(3 Pt 1):828–832. doi: 10.1083/jcb.87.3.828. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Sandvig K., Olsnes S. Entry of the toxic proteins abrin, modeccin, ricin, and diphtheria toxin into cells. I. Requirement for calcium. J Biol Chem. 1982 Jul 10;257(13):7495–7503. [PubMed] [Google Scholar]
  17. Sandvig K., Olsnes S. Entry of the toxic proteins abrin, modeccin, ricin, and diphtheria toxin into cells. II. Effect of pH, metabolic inhibitors, and ionophores and evidence for toxin penetration from endocytotic vesicles. J Biol Chem. 1982 Jul 10;257(13):7504–7513. [PubMed] [Google Scholar]
  18. Sandvig K., Olsnes S., Pihl A. Inhibitory effect of ammonium chloride and chloroquine on the entry of the toxic lectin modeccin into HeLa cells. Biochem Biophys Res Commun. 1979 Sep 27;90(2):648–655. doi: 10.1016/0006-291x(79)91284-1. [DOI] [PubMed] [Google Scholar]
  19. Stirpe F., Sandvig K., Olsnes S., Pihl A. Action of viscumin, a toxic lectin from mistletoe, on cells in culture. J Biol Chem. 1982 Nov 25;257(22):13271–13277. [PubMed] [Google Scholar]
  20. VAN HEYNINGEN W. E., GLADSTONE G. P. The neurotoxin of Shigella shigae. I. Production, purification and properties of the toxin. Br J Exp Pathol. 1953 Apr;34(2):202–216. [PMC free article] [PubMed] [Google Scholar]

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