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. 1984 May;81(9):2703–2707. doi: 10.1073/pnas.81.9.2703

Cellular ADP-ribosyltransferase with the same mechanism of action as diphtheria toxin and Pseudomonas toxin A.

H Lee, W J Iglewski
PMCID: PMC345138  PMID: 6326138

Abstract

An ADP-ribosyltransferase was found in elongation factor 2 (EF-2) preparations from polyoma virus-transformed baby hamster kidney (pyBHK) cells. Like fragment A of diphtheria toxin and Pseudomonas toxin A, this eukaryotic cellular enzyme transfers [14C]adenosine from NAD+ to EF-2. However, the cellular transferase is immunologically distinct from fragment A. The transferase also can be distinguished from fragment A and Pseudomonas toxin A by the inhibition of the activity of the former by cytoplasmic extracts and by histamine. Snake venom phosphodiesterase digestion of the [14C]adenosine-labeled EF-2 product of the cellular transferase reaction yielded [14C]AMP, indicating that the cellular enzyme is a mono(ADP-ribosyl)transferase. The forward ADP-ribosylation reaction catalyzed by the cellular enzyme is reversed by fragment A, yielding [14C]NAD+. The results strongly suggest that the cellular transferase is a mono(ADP-ribosyl)transferase, which ADP-ribosylates the same diphthamide residue of EF-2 as does fragment A and Pseudomonas toxin A.

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Selected References

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

  1. Adamietz P., Wielckens K., Bredehorst R., Lengyel H., Hilz H. Subcellular distribution of mono(ADP-ribose) protein conjugates in rat liver. Biochem Biophys Res Commun. 1981 Jul 16;101(1):96–103. doi: 10.1016/s0006-291x(81)80015-0. [DOI] [PubMed] [Google Scholar]
  2. Bredehorst R., Wielckens K., Adamietz P., Steinhagen-Thiessen E., Hilz H. Mono(ADP-ribosyl)ation and poly(ADP-ribosyl)ation of proteins in developing liver and in hepatomas: relation of conjugate subfractions to metabolic competence and proliferation rates. Eur J Biochem. 1981 Nov;120(2):267–274. doi: 10.1111/j.1432-1033.1981.tb05699.x. [DOI] [PubMed] [Google Scholar]
  3. Bredehorst R., Wielckens K., Gartemann A., Lengyel H., Klapproth K., Hilz H. Two different types of bonds linking single ADP-ribose residues covalently to proteins. Quantification in eukaryotic cells. Eur J Biochem. 1978 Dec 1;92(1):129–135. doi: 10.1111/j.1432-1033.1978.tb12730.x. [DOI] [PubMed] [Google Scholar]
  4. Brown B. A., Bodley J. W. Primary structure at the site in beef and wheat elongation factor 2 of ADP-ribosylation by diphtheria toxin. FEBS Lett. 1979 Jul 15;103(2):253–255. doi: 10.1016/0014-5793(79)81339-3. [DOI] [PubMed] [Google Scholar]
  5. Collier R. J. Diphtheria toxin: mode of action and structure. Bacteriol Rev. 1975 Mar;39(1):54–85. doi: 10.1128/br.39.1.54-85.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gill D. M., Dinius L. L. The elongation factor 2 content of mammalian cells. Assay method and relation to ribosome number. J Biol Chem. 1973 Jan 25;248(2):654–658. [PubMed] [Google Scholar]
  7. Hayaishi O., Ueda K. Poly(ADP-ribose) and ADP-ribosylation of proteins. Annu Rev Biochem. 1977;46:95–116. doi: 10.1146/annurev.bi.46.070177.000523. [DOI] [PubMed] [Google Scholar]
  8. Honjo T., Nishizuka Y., Hayaishi O. Diphtheria toxin-dependent adenosine diphosphate ribosylation of aminoacyl transferase II and inhibition of protein synthesis. J Biol Chem. 1968 Jun 25;243(12):3553–3555. [PubMed] [Google Scholar]
  9. Honjo T., Nishizuka Y., Kato I., Hayaishi O. Adenosine diphosphate ribosylation of aminoacyl transferase II and inhibition of protein synthesis by diphtheria toxin. J Biol Chem. 1971 Jul 10;246(13):4251–4260. [PubMed] [Google Scholar]
  10. Iglewski B. H., Liu P. V., Kabat D. Mechanism of action of Pseudomonas aeruginosa exotoxin Aiadenosine diphosphate-ribosylation of mammalian elongation factor 2 in vitro and in vivo. Infect Immun. 1977 Jan;15(1):138–144. doi: 10.1128/iai.15.1.138-144.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Iglewski B. H., Sadoff J., Bjorn M. J., Maxwell E. S. Pseudomonas aeruginosa exoenzyme S: an adenosine diphosphate ribosyltransferase distinct from toxin A. Proc Natl Acad Sci U S A. 1978 Jul;75(7):3211–3215. doi: 10.1073/pnas.75.7.3211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Iglewski W. J., Lee H. Purification and properties of an altered form of elongation factor 2 from mutant cells resistant to intoxication by diphtheria toxin. Eur J Biochem. 1983 Aug 1;134(2):237–240. doi: 10.1111/j.1432-1033.1983.tb07556.x. [DOI] [PubMed] [Google Scholar]
  13. Moss J., Vaughan M. Isolation of an avian erythrocyte protein possessing ADP-ribosyltransferase activity and capable of activating adenylate cyclase. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3621–3624. doi: 10.1073/pnas.75.8.3621. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Oppenheimer N. J., Bodley J. W. Diphtheria toxin. Site and configuration of ADP-ribosylation of diphthamide in elongation factor 2. J Biol Chem. 1981 Aug 25;256(16):8579–8581. [PubMed] [Google Scholar]
  15. Pappenheimer A. M., Jr Diphtheria toxin. Annu Rev Biochem. 1977;46:69–94. doi: 10.1146/annurev.bi.46.070177.000441. [DOI] [PubMed] [Google Scholar]
  16. Pappenheimer A. M., Jr, Gill D. M. Diphtheria. Science. 1973 Oct 26;182(4110):353–358. doi: 10.1126/science.182.4110.353. [DOI] [PubMed] [Google Scholar]
  17. Pappenheimer A. M., Jr, Uchida T., Harper A. A. An immunological study of the diphtheria toxin molecule. Immunochemistry. 1972 Sep;9(9):891–906. doi: 10.1016/0019-2791(72)90163-2. [DOI] [PubMed] [Google Scholar]
  18. Roberts J. H., Stard P., Giri C. P., Smulson M. Cytoplasmic poly(ADP-ribose) polymerase during the HeLa cell cycle. Arch Biochem Biophys. 1975 Nov;171(1):305–315. doi: 10.1016/0003-9861(75)90037-5. [DOI] [PubMed] [Google Scholar]
  19. Van Ness B. G., Barrowclough B., Bodley J. W. Recognition of elongation factor 2 by diphtheria toxin is not solely defined by the presence of diphthamide. FEBS Lett. 1980 Oct 20;120(1):4–6. doi: 10.1016/0014-5793(80)81032-5. [DOI] [PubMed] [Google Scholar]
  20. 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]

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