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. 1994 Nov 15;304(Pt 1):183–187. doi: 10.1042/bj3040183

Forced expression of antizyme abolishes ornithine decarboxylase activity, suppresses cellular levels of polyamines and inhibits cell growth.

Y Murakami 1, S Matsufuji 1, Y Miyazaki 1, S Hayashi 1
PMCID: PMC1137469  PMID: 7998932

Abstract

Ornithine decarboxylase (ODC) is a key enzyme in polyamine biosynthesis. It is a short-lived protein and negatively regulated by its products, polyamines. Its degradation is accelerated by the binding of antizyme, an ODC-inhibitory protein induced by polyamines. To evaluate the physiological importance of antizyme we examined the effect of forced expression of antizyme on cellular ODC and polyamine levels and cell growth. Antizyme almost completely abolished the induction of ODC by growth stimuli. This may have been caused by antizyme-induced rapid degradation of newly synthesized ODC, since the half-life of ODC complexes with antizyme was less than 5 min. Forced expression of antizyme caused reductions of cellular putrescine and spermidine levels, and inhibited cell growth, which was partially restored by the addition of putrescine. These observations suggested a critically important role of antizyme in polyamine metabolism.

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Selected References

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

  1. Auvinen M., Paasinen A., Andersson L. C., Hölttä E. Ornithine decarboxylase activity is critical for cell transformation. Nature. 1992 Nov 26;360(6402):355–358. doi: 10.1038/360355a0. [DOI] [PubMed] [Google Scholar]
  2. Choi J. H., Scheffler I. E. Chinese hamster ovary cells resistant to alpha-difluoromethylornithine are overproducers of ornithine decarboxylase. J Biol Chem. 1983 Oct 25;258(20):12601–12608. [PubMed] [Google Scholar]
  3. Davis R. H., Morris D. R., Coffino P. Sequestered end products and enzyme regulation: the case of ornithine decarboxylase. Microbiol Rev. 1992 Jun;56(2):280–290. doi: 10.1128/mr.56.2.280-290.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dircks L., Grens A., Slezynger T. C., Scheffler I. E. Posttranscriptional regulation of ornithine decarboxylase activity. J Cell Physiol. 1986 Mar;126(3):371–378. doi: 10.1002/jcp.1041260307. [DOI] [PubMed] [Google Scholar]
  5. Fong W. F., Heller J. S., Canellakis E. S. The appearance of an ornithine decarboxylase inhibitory protein upon the addition of putrescine to cell cultures. Biochim Biophys Acta. 1976 Apr 23;428(2):456–465. doi: 10.1016/0304-4165(76)90054-4. [DOI] [PubMed] [Google Scholar]
  6. Gupta M., Coffino P. Mouse ornithine decarboxylase. Complete amino acid sequence deduced from cDNA. J Biol Chem. 1985 Mar 10;260(5):2941–2944. [PubMed] [Google Scholar]
  7. Heby O. Role of polyamines in the control of cell proliferation and differentiation. Differentiation. 1981;19(1):1–20. doi: 10.1111/j.1432-0436.1981.tb01123.x. [DOI] [PubMed] [Google Scholar]
  8. Heller J. S., Fong W. F., Canellakis E. S. Induction of a protein inhibitor to ornithine decarboxylase by the end products of its reaction. Proc Natl Acad Sci U S A. 1976 Jun;73(6):1858–1862. doi: 10.1073/pnas.73.6.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hershko A., Ciechanover A. The ubiquitin system for protein degradation. Annu Rev Biochem. 1992;61:761–807. doi: 10.1146/annurev.bi.61.070192.003553. [DOI] [PubMed] [Google Scholar]
  10. Hölttä E., Pohjanpelto P. Control of ornithine decarboxylase in Chinese hamster ovary cells by polyamines. Translational inhibition of synthesis and acceleration of degradation of the enzyme by putrescine, spermidine, and spermine. J Biol Chem. 1986 Jul 15;261(20):9502–9508. [PubMed] [Google Scholar]
  11. Ito K., Kashiwagi K., Watanabe S., Kameji T., Hayashi S., Igarashi K. Influence of the 5'-untranslated region of ornithine decarboxylase mRNA and spermidine on ornithine decarboxylase synthesis. J Biol Chem. 1990 Aug 5;265(22):13036–13041. [PubMed] [Google Scholar]
  12. Kaczmarek L., Calabretta B., Ferrari S., de Riel J. K. Cell-cycle-dependent expression of human ornithine decarboxylase. J Cell Physiol. 1987 Sep;132(3):545–551. doi: 10.1002/jcp.1041320318. [DOI] [PubMed] [Google Scholar]
  13. Kanamoto R., Kameji T., Iwashita S., Igarashi K., Hayashi S. Spermidine-induced destabilization of ornithine decarboxylase (ODC) is mediated by accumulation of antizyme in ODC-overproducing variant cells. J Biol Chem. 1993 May 5;268(13):9393–9399. [PubMed] [Google Scholar]
  14. Kanamoto R., Utsunomiya K., Kameji T., Hayashi S. Effects of putrescine on synthesis and degradation of ornithine decarboxylase in primary cultured hepatocytes. Eur J Biochem. 1986 Feb 3;154(3):539–544. doi: 10.1111/j.1432-1033.1986.tb09432.x. [DOI] [PubMed] [Google Scholar]
  15. Kitani T., Fujisawa H. Purification and some properties of a protein inhibitor (antizyme) of ornithine decarboxylase from rat liver. J Biol Chem. 1984 Aug 25;259(16):10036–10040. [PubMed] [Google Scholar]
  16. Li X., Coffino P. Degradation of ornithine decarboxylase: exposure of the C-terminal target by a polyamine-inducible inhibitory protein. Mol Cell Biol. 1993 Apr;13(4):2377–2383. doi: 10.1128/mcb.13.4.2377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Li X., Coffino P. Regulated degradation of ornithine decarboxylase requires interaction with the polyamine-inducible protein antizyme. Mol Cell Biol. 1992 Aug;12(8):3556–3562. doi: 10.1128/mcb.12.8.3556. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matsufuji S., Miyazaki Y., Kanamoto R., Kameji T., Murakami Y., Baby T. G., Fujita K., Ohno T., Hayashi S. Analyses of ornithine decarboxylase antizyme mRNA with a cDNA cloned from rat liver. J Biochem. 1990 Sep;108(3):365–371. doi: 10.1093/oxfordjournals.jbchem.a123207. [DOI] [PubMed] [Google Scholar]
  19. McConlogue L., Dana S. L., Coffino P. Multiple mechanisms are responsible for altered expression of ornithine decarboxylase in overproducing variant cells. Mol Cell Biol. 1986 Aug;6(8):2865–2871. doi: 10.1128/mcb.6.8.2865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mitchell J. L., Chen H. J. Conformational changes in ornithine decarboxylase enable recognition by antizyme. Biochim Biophys Acta. 1990 Jan 19;1037(1):115–121. doi: 10.1016/0167-4838(90)90109-s. [DOI] [PubMed] [Google Scholar]
  21. Mitchell J. L., Diveley R. R., Jr, Bareyal-Leyser A. Feedback repression of polyamine uptake into mammalian cells requires active protein synthesis. Biochem Biophys Res Commun. 1992 Jul 15;186(1):81–88. doi: 10.1016/s0006-291x(05)80778-8. [DOI] [PubMed] [Google Scholar]
  22. Murakami Y., Marumo M., Hayashi S. Ornithine decarboxylase antizyme in kidneys of male and female mice. Biochem J. 1988 Sep 1;254(2):367–372. doi: 10.1042/bj2540367. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Murakami Y., Matsufuji S., Kameji T., Hayashi S., Igarashi K., Tamura T., Tanaka K., Ichihara A. Ornithine decarboxylase is degraded by the 26S proteasome without ubiquitination. Nature. 1992 Dec 10;360(6404):597–599. doi: 10.1038/360597a0. [DOI] [PubMed] [Google Scholar]
  24. Murakami Y., Matsufuji S., Miyazaki Y., Hayashi S. Destabilization of ornithine decarboxylase by transfected antizyme gene expression in hepatoma tissue culture cells. J Biol Chem. 1992 Jul 5;267(19):13138–13141. [PubMed] [Google Scholar]
  25. Murakami Y., Tanaka K., Matsufuji S., Miyazaki Y., Hayashi S. Antizyme, a protein induced by polyamines, accelerates the degradation of ornithine decarboxylase in Chinese-hamster ovary-cell extracts. Biochem J. 1992 May 1;283(Pt 3):661–664. doi: 10.1042/bj2830661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pegg A. E. Polyamine metabolism and its importance in neoplastic growth and a target for chemotherapy. Cancer Res. 1988 Feb 15;48(4):759–774. [PubMed] [Google Scholar]
  27. Pegg A. E. Recent advances in the biochemistry of polyamines in eukaryotes. Biochem J. 1986 Mar 1;234(2):249–262. doi: 10.1042/bj2340249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Persson L., Holm I., Heby O. Translational regulation of ornithine decarboxylase by polyamines. FEBS Lett. 1986 Sep 15;205(2):175–178. doi: 10.1016/0014-5793(86)80892-4. [DOI] [PubMed] [Google Scholar]
  29. Prouty W. F. Ornithine decarboxylase inactivation in HeLa cells. J Cell Physiol. 1976 Sep;89(1):65–76. doi: 10.1002/jcp.1040890107. [DOI] [PubMed] [Google Scholar]
  30. Russell D. H., Snyder S. H. Amine synthesis in regenerating rat liver: extremely rapid turnover of ornithine decarboxylase. Mol Pharmacol. 1969 May;5(3):253–262. [PubMed] [Google Scholar]
  31. Steglich C., Scheffler I. E. An ornithine decarboxylase-deficient mutant of Chinese hamster ovary cells. J Biol Chem. 1982 Apr 25;257(8):4603–4609. [PubMed] [Google Scholar]
  32. Van Steeg H., Van Oostrom C. T., Hodemaekers H. M., Peters L., Thomas A. A. The translation in vitro of rat ornithine decarboxylase mRNA is blocked by its 5' untranslated region in a polyamine-independent way. Biochem J. 1991 Mar 1;274(Pt 2):521–526. doi: 10.1042/bj2740521. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. van Daalen Wetters T., Macrae M., Brabant M., Sittler A., Coffino P. Polyamine-mediated regulation of mouse ornithine decarboxylase is posttranslational. Mol Cell Biol. 1989 Dec;9(12):5484–5490. doi: 10.1128/mcb.9.12.5484. [DOI] [PMC free article] [PubMed] [Google Scholar]

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