Advertisement

Post-Translational Modifications of Eukaryotic Initiation Factor-5A (eIF-5a) as a New Target for Anti-Cancer Therapy

  • Michele Caraglia
  • Pierosandro Tagliaferri
  • Alfredo Budillon
  • Alberto Abbruzzese
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 472)

Abstract

Eukaryotic translation initiation factor 5A (eIF-5A) is the only cell protein that contains the unusual basic amino acid hypusine [Nε-(4-amino-2-hydroxybutyl)lysine]. Hypusine is formed by the transfer of the butylamine portion from spermidine to the ε-amino group of a specific lysine residue of eIF-5A precursor and the subsequent hydroxylation at carbon 2 of the incoming 4-aminobutyl moiety. Agents that reduce cell hypusine levels inhibit the growth of mammalian cells. These observations suggest that hypusine is crucial for proliferation and transformation of eukaryotic cells. Here we have studied whether the inhibition of hypusine synthesis can potentiate the anticancer activity of the anti-tumour agents interferon-α (IFNα) and cytosine arabinoside (ara-C). We have found that IFNα increased epidermal growth factor receptor (EGF-R) expression, but reduced S phase and proliferative marker expression in human epidermoid KB cells and that this effect was antagonised by epidermal growth factor (EGF). Growth inhibition induced by IFNα was paralleled by decreased hypusine synthesis and, when EGF counteracted anti-proliferative effects, a reconstitution of hypusine levels was recorded. We also studied the effects of IFNα on the cytotoxicity of the recombinant toxin TP40 which inhibits elongation factor 2, another step of protein synthesis, through EGF-R binding and internalisation; IFNα induced an about 27-fold increase of TP40 cytotoxicity in KB cells.

Ara-C, another antineoplastic agent commonly used in haematologic malignancies, induced both apoptosis and iron depletion in human acute myeloid leukaemic cells. The combination of ara-C and of the iron chelator desferioxamine, a strong inhibitor of hypusine synthesis, had a synergistic activity on apoptosis in these cells. The data strongly suggest that the post-translational modifications of eIF-5A could be a suitable target for the potentiation of the activity of anti-cancer agents.

Keywords

Epidermal Growth Factor Receptor High Performance Liquid Chromatography Epidermal Growth Factor Receptor Expression Haem Iron Cytosine Arabinoside 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Milovic, V., Deubner, C., Zeuzem, S., Piiper, A., Caspary, W.F., and Stein, J. (1995) EGF stimulates polyamine uptake in Caco-2 cells. Biochem. Biophys. Res. Commun. 206, 962–968.PubMedCrossRefGoogle Scholar
  2. 2.
    Wojciechowski, K., Trzeciak, L., Konturek, S.J., and Ostrowski (1995) Inhibition of acid secretory response and induction of ornithine decarboxylase and its mRNA by TGF alpha and EGF in isolated rat gastric glands. J. Regul. Pept. 56, 1–8.CrossRefGoogle Scholar
  3. 3.
    Panagiotidis, C., Artandi, S., Calame, K., and Silverstein, S.J. (1995) Polyamines alter sequence-specific DNA-protein interactions. Nucleic Acid Res. 23, 1800–1809.PubMedCrossRefGoogle Scholar
  4. 4.
    Cooper, H.L., Park, M.H., Folk, J.E., Safer, B., and Braverman, R. (1983) Identification of the hypusine-containing protein hy+ as translation initiation factor eIF-4D. Proc. Natl. Acad. Sci. USA 80, 1854–1857.PubMedCrossRefGoogle Scholar
  5. 5.
    Park, M.H., Wolff, E.C., and Folk, J.E. (1993) Hypusine: its post-translational formation in eukaryotic initiation factor 5A and its potential role in cellular regulation. BioFactors 4, 95–104.PubMedGoogle Scholar
  6. 6.
    Park, M.H. and Wolff, E.C. (1988) Cell-free synthesis of deoxyhypusine. Separation of protein substrate and enzyme and identification of 1,3-diaminopropane as a product of spermidine cleavage. J. Biol. Chem. 263, 15264–15269.PubMedGoogle Scholar
  7. 7.
    Wolff, E.C., Park, M.H., and Folk, J.E. (1990) Cleavage of spermidine as the first step in deoxyhypusine synthesis. The role of NAD. J. Biol. Chem. 265, 4793–4799.PubMedGoogle Scholar
  8. 8.
    Park, M.H., Chung, S.I., Cooper, H.L., and Folk, J.E. (1984) The mammalian hypusine-containing protein, eukaryotic initiation factor 4D. J. Biol. Chem. 259, 4563–4565.PubMedGoogle Scholar
  9. 9.
    Beninati, S., Abbruzzese A., and Folk J.E. (1990) High-performance liquid chromatographic method for determination of hypusine and deoxyhypusine. Anal. Biochem. 184, 16–20.PubMedCrossRefGoogle Scholar
  10. 10.
    Park, M.H., Cooper, H.L., and Folk, J.E. (1982) The biosynthesis of protein-bound hypusine amino-2-hydroxybutyl)lysine): lysine as the amino acid precursor and the intermediate role of deoxhypusine (N`-(4-amino-butyl)lysine). J. Biol. Chem. 257, 7217–7222.PubMedGoogle Scholar
  11. 11.
    Abbruzzese, A., Park, M.H., and Folk, J.E. (1986) Deoxhypusine hydroxylase from rat testis. Partial purification and characterization. J. Biol. Chem. 261, 3085–3089.PubMedGoogle Scholar
  12. 12.
    Abbruzzese, A., Park, M.H., and Folk, J.E. (1985) Deoxyhypusine hydroxylase: distribution and partial purification from rat testis. Fed. Proc. 44, 1487.Google Scholar
  13. 13.
    Beninati, S., Nicolini, L., Jakus, J., Passeggio, A., and Abbruzzese, A. (1995) Identification of a substrate site for transglutaminases on the human protein synthesis initiation factor 5A. Biochem. J. 305, 725–728.PubMedGoogle Scholar
  14. 14.
    Beninati, S., Gentile, V., Caraglia, M., Lentini, A., Tagliaferri, E, and Abbruzzese, A. (1998) Tissue transglutaminase expression affects hypusine metabolism in balb-C 3T3 cells. FEBS Letters 437, 34–38.PubMedCrossRefGoogle Scholar
  15. 15.
    Hershey, J.W.B. (1991) Translational control in mammalian cells. Annu. Rev. Biochem. 61, 717–755.CrossRefGoogle Scholar
  16. 16.
    Stiuso, R, Colonna, G., Ragone, R., Caraglia, M., Hershey, J.W.B., Beninati, S., and Abbruzzese A. (1999) Structural organization of the human eukaryotic initiation factor 5a precursor and its site-directed variant lys 50- arg. Amino Acids 16, 91–106.PubMedCrossRefGoogle Scholar
  17. 17.
    Park, M.H., Wolff, E.C., Smit-McBride, Z., Hershey, J.W.B., and Folk, J.E. (1991) Comparison of the activities of variant forms of eIF-4D: the requirement for hypusine or deoxyhypusine. J. Biol. Chem. 266, 7988–7994.PubMedGoogle Scholar
  18. 18.
    Cooper, H.L., Park, M.H., and Folk, J.E. (1982) Posttranslational formation of hypusine in a single major protein occurs generally in growing cells and is associated with activation of lymphocyte growth. Cell 29, 791–797.PubMedCrossRefGoogle Scholar
  19. 19.
    Lentini, A., Mattioli, R, Nicolini, L., Pietrini, A., Abbruzzese, A., and Beninati, S. (1997) Anti-invasive effects of theophylline on experimental B16–F10 melanoma lung metastasis. Cancer J. 10, 274–278.Google Scholar
  20. 20.
    Abbruzzese, A. (1988) Developmental pattern for deoxyhypusine hydroxylase in rat brain. J. Neurochem. 50, 695–699.PubMedCrossRefGoogle Scholar
  21. 21.
    Abbruzzese, A., Park, M.H., Beninati, S., and Folk, J.E. (1989) Inhibition of deoxyhypusine hydroxylase by polyamines and by a deoxyhypusine peptide. Biochem. Biophys. Acta 997, 248–255.PubMedCrossRefGoogle Scholar
  22. 22.
    Abbruzzese, A., Hanauske-Abel, H.M., Park, M.H., Henke, S., and Folk, J.E. (1991) The active site of deoxyhypusyl hydroxylase: use of catecholpeptides and their component chelator and peptide moieties as molecular probes. Biochem. Biophys. Acta 1077, 159–166.Google Scholar
  23. 23.
    Jakus, J., Wolff, E.C., Park, M.H., and Folk, J.E. (1993) Features of the spermidine-binding site of deoxyhypusine synthase as derived from inhibition studies. J. Biol. Chem. 268, 13151–13159.PubMedGoogle Scholar
  24. 24.
    Park, M.H., Wolff, E.C., and Folk, J.E. (1993) Is hypusine essential for eukaryotic cell proliferation? Trends Biochem. Sci. 18, 475–479.PubMedCrossRefGoogle Scholar
  25. 25.
    Budillon, A., Tagliaferri, R, Caraglia, M., Torrisi, M.R., Normanno, N., Iacobelli, S., Palmieri, G., Stoppelli, M.P., Frati, L., and Bianco, A.R. (1991) Upregulation of epidermal growth factor receptor induced by alpha-interferon in human epidermoid cancer cells. Cancer Res. 51, 1294–1299.PubMedGoogle Scholar
  26. 26.
    Caraglia, M., Leardi, A., Corradino, S., Ciardiello, E, Budillon, A., Guarrasi, R., Bianco, A.R., and Tagliaferri, P. (1995) “a-Interferon potentiates epidermal growth factor receptor-mediated effects on human epidermoid carcinoma KB cells. Int. J. Cancer, 61, 342–347.PubMedCrossRefGoogle Scholar
  27. 27.
    Caraglia, M.,Tagliaferri, R, Correale, R, Genua, G., Pinto, A., Del Vecchio, S., Esposito, G., and Bianco, A.R. (1993) Cytosine arabinoside increases the binding of [1251]-labelled epidermal growth factor and [125I]-transferrin and enhances the in vitro targeting of human tumor cells with anti-(growth factor receptor) mAb. Cancer Immunol. Immunother. 37, 150–156.PubMedCrossRefGoogle Scholar
  28. 28.
    Caraglia, M., Leardi, A., Improta, S., Perin, V., Ricciardi, B., Arra, C., Ferraro, R, Fabbrocini, A., Pinto, A., Bianco, A.R., and Tagliaferri, P. (1997) Transient exposure to cytarabine increases peptide growth factor receptor expression and tumorigenicity of melanoma cells. Anticancer Res. 17, 2369–2376.PubMedGoogle Scholar
  29. 29.
    Tagliaferri, R, Caraglia, M., Muraro, R., Budillon, A., Pinto, A., and Bianco, A.R. (1994) Pharmacological modulation of peptide growth factor receptor expression on tumor cells as a basis for cancer therapy. Anti-Cancer Drugs 5, 379–393.PubMedCrossRefGoogle Scholar
  30. 30.
    Pai, L.H., Gallo, M.G., Fitzgerald, D.J., and Pastan, I. (1991) Antitumor activity of a transforming growth factor alpha-Pseudomonas exotoxin fusion protein (TGF-alpha-PE40). Cancer Res. 51, 2808–2812.PubMedGoogle Scholar
  31. 31.
    Caraglia, M., Libroia, A.M., Corradino, S., Coppola, V., Guarrasi, R., Barile, C., Genua, G., Bianco, A.R., and Tagliaferri, P. (1994) “a-Interferon induces depletion of intracellular iron content and upregulation of functional transferrin receptors on human epidermoid cancer KB cells. Biochem. Biophy. Res. Commun. 203, 281–288.CrossRefGoogle Scholar
  32. 32.
    Park, M.H. (1987) Regulation of biosynthesis of hypusine in Chinese hamster ovary cells. Evidence for eIF-4D precursor polypeptides. J. Biol. Chem. 262, 12730–12734.PubMedGoogle Scholar
  33. 33.
    Lalande, M. and Hanausske-Abel, H.M. (1990) A new compound which reversibly arrests T lymphocyte cell cycle near the Gl/S boundary. Exp. Cell. Res. 188, 117–121.PubMedCrossRefGoogle Scholar
  34. 34.
    Park, M.H., Cooper, H.L., and Folk, J.E. (1981) Identification of hypusine, an unusual amino acid, in a protein from human lymphocytes and of spermidine as its biosynthetic precursor. Proc. Natl. Acad. Sci. USA 78, 2869–2873.PubMedCrossRefGoogle Scholar
  35. 35.
    Russell, D.H. and Dude, B.G. (1978) Polyamines as Biochemical Markers of Normal and Malignant Growth, 1–178, Raven Press, NY.Google Scholar
  36. 36.
    Lentini, A., Kleinman, H.K., Mattioli, E, Autuori, V., Nicolini, L., Pietrini, A., Abbruzzese, A., Cardinale, M., and Beninati, S. (1998) Inhibition of melanoma pulmonary metastasis by methylxanthines due to decreased invasion and proliferation. Melanoma Res. 8, 131–137.PubMedCrossRefGoogle Scholar
  37. 37.
    Beninati, S., Abbruzzese, A., and Cardinale, M. (1993) Differences in the posttranslational modification of proteins by polyamines between weakly and highly metastatic B16 melanoma cells. Int. J. Cancer 53, 792–797.PubMedCrossRefGoogle Scholar
  38. 38.
    Siegall, C.B., Xu, Y.H., Chaudary, V.K., Adhya, S., Fitzgerald, D., and Pastan I. (1989) Cytotoxic activities of a fusion protein comprised of TGF alpha and Pseudomonas exotoxin. FASEB J. 3, 2647–2652.PubMedGoogle Scholar
  39. 39.
    Theil, E.C. (1990) Regulation of ferritin and transferrin receptor in mRNAs. J. Biol. Chem. 265, 4771–4774.PubMedGoogle Scholar
  40. 40.
    Estrov, Z., Tawa, A., Wang, X-H., Dubé, I.D., Sulh, H., Cohen, A., Gelfand, E.W., and Freedman, M.H. (1987) In vitro and in vivo effects of deferoxamine in neonatal acute leukemia. Blood 69, 757–761.PubMedGoogle Scholar
  41. 41.
    Leardi, A., Caraglia, M., Selleri, C., Pepe, S., Pizzi, C., Notaro, R., Fabbrocini, A., Musicô, M., Abbruzzese, A., Bianco, A.R., and Tagliaferri, P. (1998) desferioxamine increases iron depletion and apoptosis induced by ara-c on human myeloid leukemic cells. Br. J. Haematol. 102, 746–752.PubMedCrossRefGoogle Scholar
  42. 42.
    Caraglia, M., Passeggio, A., Beninati, S., Leardi, A., Nicolini, L., Improta, S., Pinto, A., Bianco, A.R., Tagliaferri, P., and Abbruzzese, A. (1997) Interferon a2 recombinant and epidermal growth factor modulate proliferation and hypusine synthesis in human epidermoid cancer KB cells. Biochem. J. 324, 737–741.PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1999

Authors and Affiliations

  • Michele Caraglia
    • 1
  • Pierosandro Tagliaferri
    • 2
  • Alfredo Budillon
    • 3
  • Alberto Abbruzzese
    • 1
  1. 1.Department of Biochemistry and Biophysics “F. Cedrangolo”Second University of NaplesItaly
  2. 2.Department of Endocrinology and Molecular and Clinical OncologyUniversity “Federico II” of NaplesItaly
  3. 3.Division of Experimental Oncology CNational Tumour Institute Foundation “G. Pascale,”NaplesItaly

Personalised recommendations