Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1982 May;2(5):588–592. doi: 10.1128/mcb.2.5.588

Saccharomyces cerevisiae spheroplasts are sensitive to the action of diphtheria toxin.

S Murakami, J W Bodley, D M Livingston
PMCID: PMC369829  PMID: 7050676

Abstract

Diphtheria toxin kills spheroplasts of Saccharomyces cerevisiae but not the intact yeast cells. After 2 h of exposure to ca. 10(-7) M toxin, less than 1% of spheroplasts were able to regenerate into intact cells. The same high levels of toxin inhibited the rate of protein synthesis by more than 90% within 1 h, whereas RNA and DNA synthesis were not inhibited until 4 h or exposure. Both killing and protein synthesis inhibition were dependent on toxin concentration. The nature of the toxin-cell interaction was also studied by using fragments of intact toxin and mutant toxin proteins. Neither toxin fragment A nor CRM45 nor CRM197 affected spheroplasts, but CRM197 and ATP prevented the inhibitory action of intact toxin. These results suggest that toxin acts on S. cerevisiae spheroplasts in much the same manner as it acts on sensitive mammalian cells.

Full text

PDF
588

Selected References

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

  1. Beggs J. D. Transformation of yeast by a replicating hybrid plasmid. Nature. 1978 Sep 14;275(5676):104–109. doi: 10.1038/275104a0. [DOI] [PubMed] [Google Scholar]
  2. Boquet P., Pappenheimer A. M., Jr Interaction of diphtheria toxin with mammalian cell membranes. J Biol Chem. 1976 Sep 25;251(18):5770–5778. [PubMed] [Google Scholar]
  3. Collier R. J., Kandel J. Structure and activity of diphtheria toxin. I. Thiol-dependent dissociation of a fraction of toxin into enzymically active and inactive fragments. J Biol Chem. 1971 Mar 10;246(5):1496–1503. [PubMed] [Google Scholar]
  4. Draper R. K., Chin D., Eurey-Owens D., Scheffler I. E., Simon M. I. Biochemical and genetic characterization of three hamster cell mutants resistant to diphtheria toxin. J Cell Biol. 1979 Oct;83(1):116–125. doi: 10.1083/jcb.83.1.116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Drazin R., Kandel J., Collier R. J. Structure and activity of diphtheria toxin. II. Attack by trypsin at a specific site within the intact toxin molecule. J Biol Chem. 1971 Mar 10;246(5):1504–1510. [PubMed] [Google Scholar]
  6. Gill D. M., Dinius L. L. Observations on the structure of diphtheria toxin. J Biol Chem. 1971 Mar 10;246(5):1485–1491. [PubMed] [Google Scholar]
  7. Gill D. M., Pappenheimer A. M., Jr Structure-activity relationships in diphtheria toxin. J Biol Chem. 1971 Mar 10;246(5):1492–1495. [PubMed] [Google Scholar]
  8. Gupta R. S., Siminovitch L. Diphtheria-toxin-resistant mutants of CHO cells affected in protein synthesis: a novel phenotype. Somatic Cell Genet. 1978 Sep;4(5):553–571. doi: 10.1007/BF01542926. [DOI] [PubMed] [Google Scholar]
  9. Gupta R. S., Siminovitch L. Isolation and characterization of mutants of human diploid fibroblasts resistant to diphtheria toxin. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3337–3340. doi: 10.1073/pnas.75.7.3337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hartwell L. H. Macromolecule synthesis in temperature-sensitive mutants of yeast. J Bacteriol. 1967 May;93(5):1662–1670. doi: 10.1128/jb.93.5.1662-1670.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Middlebrook J. L., Dorland R. B., Leppla S. H. Association of diphtheria toxin with Vero cells. Demonstration of a receptor. J Biol Chem. 1978 Oct 25;253(20):7325–7330. [PubMed] [Google Scholar]
  12. Middlebrook J. L., Dorland R. B. Protection of mammalian cells from diphtheria toxin by exogenous nucleotides. Can J Microbiol. 1979 Mar;25(3):285–290. doi: 10.1139/m79-046. [DOI] [PubMed] [Google Scholar]
  13. Middlebrook J. L., Dorland R. B. Response of cultured mammalian cells to the exotoxins of Pseudomonas aeruginosa and Corynebacterium diphtheriae: differential cytotoxicity. Can J Microbiol. 1977 Feb;23(2):183–189. doi: 10.1139/m77-026. [DOI] [PubMed] [Google Scholar]
  14. Moehring J. M., Moehring T. J. Characterization of the diphtheria toxin-resistance system in Chinese hamster ovary cells. Somatic Cell Genet. 1979 Jul;5(4):453–468. doi: 10.1007/BF01538880. [DOI] [PubMed] [Google Scholar]
  15. Moehring J. M., Moehring T. J., Danley D. E. Posttranslational modification of elongation factor 2 in diphtheria-toxin-resistant mutants of CHO-K1 cells. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1010–1014. doi: 10.1073/pnas.77.2.1010. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moehring T. J., Danley D. E., Moehring J. M. Codominant translational mutants of Chinese hamster ovary cells selected with diphtheria toxin. Somatic Cell Genet. 1979 Jul;5(4):469–480. doi: 10.1007/BF01538881. [DOI] [PubMed] [Google Scholar]
  17. Moehring T. J., Moehring J. M. Selection and characterization of cells resistant to diphtheria toxin and pseudomonas exotoxin A: presumptive translational mutants. Cell. 1977 Jun;11(2):447–454. doi: 10.1016/0092-8674(77)90063-0. [DOI] [PubMed] [Google Scholar]
  18. Uchida T., Pappenheimer A. M., Jr, Greany R. Diphtheria toxin and related proteins. I. Isolation and properties of mutant proteins serologically related to diphtheria toxin. J Biol Chem. 1973 Jun 10;248(11):3838–3844. [PubMed] [Google Scholar]
  19. Uchida T., Pappenheimer A. M., Jr, Harper A. A. Diphtheria toxin and related proteins. 3. Reconstitution of hybrid "diphtheria toxin" from nontoxic mutant proteins. J Biol Chem. 1973 Jun 10;248(11):3851–3854. [PubMed] [Google Scholar]
  20. Uchida T., Pappenheimer A. M., Jr, Harper A. A. Diphtheria toxin and related proteins. II. Kinetic studies on intoxication of HeLa cells by diphtheria toxin and related proteins. J Biol Chem. 1973 Jun 10;248(11):3845–3850. [PubMed] [Google Scholar]
  21. Van Ness B. G., Howard J. B., Bodley J. W. ADP-ribosylation of elongation factor 2 by diphtheria toxin. Isolation and properties of the novel ribosyl-amino acid and its hydrolysis products. J Biol Chem. 1980 Nov 25;255(22):10717–10720. [PubMed] [Google Scholar]
  22. Van Ness B. G., Howard J. B., Bodley J. W. ADP-ribosylation of elongation factor 2 by diphtheria toxin. NMR spectra and proposed structures of ribosyl-diphthamide and its hydrolysis products. J Biol Chem. 1980 Nov 25;255(22):10710–10716. [PubMed] [Google Scholar]
  23. Van Ness B. G., Howard J. B., Bodley J. W. Isolation and properties of the trypsin-derived ADP-ribosyl peptide from diphtheria toxin-modified yeast elongation factor 2. J Biol Chem. 1978 Dec 25;253(24):8687–8690. [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES