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. 1998 Feb 1;329(Pt 3):453–459. doi: 10.1042/bj3290453

Osmotic stress induces variation in cellular levels of ornithine decarboxylase-antizyme.

J L Mitchell 1, G G Judd 1, A Leyser 1, C Choe 1
PMCID: PMC1219064  PMID: 9445370

Abstract

The polyamines, and especially putrescine, play an integral role in the physiological response of cells to varying extracellular osmotic conditions. Ornithine decarboxylase (ODC) synthesis and stability, as well as the activity of the polyamine transporter, had all been reported to be very sensitive to media osmolarity in different cells and tissues, yet the mechanism of this complex, co-ordinated response was not known. In this study we have determined that all these aspects of osmotic-shock response may be mediated by the common regulatory protein, ODC-antizyme. HTC cells were induced for antizyme and then exposed to media of reduced osmotic strength. Both antizyme activity and protein decreased rapidly, under these conditions, to new steady-state levels that depended upon the degree of reduction in media tonicity. This antizyme reduction was found to be due to a rapid increase in antizyme degradation, with a half-life decrease from 75 min down to 45 min occurring immediately upon exchanging media. In complementary experiments, increased media tonicity induced elevated antizyme levels and stability. The sensitivity of antizyme turnover to osmotic conditions was also observed in DH23b cells, which contain elevated levels of more stable antizyme. Interestingly, the two main antizyme proteins, AZ-1 and AZ-2 (presumably products from the first and second translational start sites), differed in their responses to these changing osmotic conditions. Just as feedback regulation of antizyme synthesis provides an effective mechanism for maintaining stable polyamine levels, these studies suggest that alteration in the rate of antizyme degradation may be the mechanism whereby cells adjust steady-state polyamine levels in response to stimulation or stress.

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Selected References

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  1. Blagbrough I. S., Brackley P. T., Bruce M., Bycroft B. W., Mather A. J., Millington S., Sudan H. L., Usherwood P. N. Arthropod toxins as leads for novel insecticides: an assessment of polyamine amides as glutamate antagonists. Toxicon. 1992 Mar;30(3):303–322. doi: 10.1016/0041-0101(92)90871-2. [DOI] [PubMed] [Google Scholar]
  2. Fontana L., Colombatto S., Grillo M. A. Regulation of spermidine transport in L1210 cells. Int J Biochem. 1993 Oct;25(10):1497–1500. doi: 10.1016/0020-711x(93)90696-c. [DOI] [PubMed] [Google Scholar]
  3. Friedman Y., Park S., Levasseur S., Burke G. Activation of thyroid ornithine decarboxylase (ODC) in vitro by hypotonicity; a possible mechanism for ODC induction. Biochem Biophys Res Commun. 1977 Jul 11;77(1):57–64. doi: 10.1016/s0006-291x(77)80164-2. [DOI] [PubMed] [Google Scholar]
  4. Hayashi S., Murakami Y., Matsufuji S. Ornithine decarboxylase antizyme: a novel type of regulatory protein. Trends Biochem Sci. 1996 Jan;21(1):27–30. [PubMed] [Google Scholar]
  5. Hayashi S., Murakami Y. Rapid and regulated degradation of ornithine decarboxylase. Biochem J. 1995 Feb 15;306(Pt 1):1–10. doi: 10.1042/bj3060001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Huang L. E., Caruccio L., Liu A. Y., Chen K. Y. Rapid activation of the heat shock transcription factor, HSF1, by hypo-osmotic stress in mammalian cells. Biochem J. 1995 Apr 15;307(Pt 2):347–352. doi: 10.1042/bj3070347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Käpyaho K., Jänne J. Regulation of putrescine metabolism in Ehrlich ascites carcinoma cells exposed to hypotonic medium. Biochim Biophys Acta. 1982 Jan 12;714(1):93–100. [PubMed] [Google Scholar]
  8. Lundgren D. W. Effect of hypotonic stress on ornithine decarboxylase mRNA expression in cultured cells. J Biol Chem. 1992 Apr 5;267(10):6841–6847. [PubMed] [Google Scholar]
  9. Lövkvist-Wallström E., Stjernborg-Ulvsbäck L., Scheffler I. E., Persson L. Regulation of mammalian ornithine decarboxylase. Studies on the induction of the enzyme by hypotonic stress. Eur J Biochem. 1995 Jul 1;231(1):40–44. [PubMed] [Google Scholar]
  10. Mamroud-Kidron E., Omer-Itsicovich M., Bercovich Z., Tobias K. E., Rom E., Kahana C. A unified pathway for the degradation of ornithine decarboxylase in reticulocyte lysate requires interaction with the polyamine-induced protein, ornithine decarboxylase antizyme. Eur J Biochem. 1994 Dec 1;226(2):547–554. doi: 10.1111/j.1432-1033.1994.tb20079.x. [DOI] [PubMed] [Google Scholar]
  11. Matsufuji S., Matsufuji T., Miyazaki Y., Murakami Y., Atkins J. F., Gesteland R. F., Hayashi S. Autoregulatory frameshifting in decoding mammalian ornithine decarboxylase antizyme. Cell. 1995 Jan 13;80(1):51–60. doi: 10.1016/0092-8674(95)90450-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. Mitchell J. L., Choe C. Y., Judd G. G., Daghfal D. J., Kurzeja R. J., Leyser A. Overproduction of stable ornithine decarboxylase and antizyme in the difluoromethylornithine-resistant cell line DH23b. Biochem J. 1996 Aug 1;317(Pt 3):811–816. doi: 10.1042/bj3170811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mitchell J. L., Choe C. Y., Judd G. G. Feedback repression of ornithine decarboxylase synthesis mediated by antizyme. Biochem J. 1996 Dec 15;320(Pt 3):755–760. doi: 10.1042/bj3200755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Mitchell J. L., Hoff J. A., Bareyal-Leyser A. Stable ornithine decarboxylase in a rat hepatoma cell line selected for resistance to alpha-difluoromethylornithine. Arch Biochem Biophys. 1991 Oct;290(1):143–152. doi: 10.1016/0003-9861(91)90600-n. [DOI] [PubMed] [Google Scholar]
  18. Mitchell J. L., Judd G. G., Bareyal-Leyser A., Ling S. Y. Feedback repression of polyamine transport is mediated by antizyme in mammalian tissue-culture cells. Biochem J. 1994 Apr 1;299(Pt 1):19–22. doi: 10.1042/bj2990019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Munro G. F., Hercules K., Morgan J., Sauerbier W. Dependence of the putrescine content of Escherichia coli on the osmotic strength of the medium. J Biol Chem. 1972 Feb 25;247(4):1272–1280. [PubMed] [Google Scholar]
  20. Munro G. F., Miller R. A., Bell C. A., Verderber E. L. Effects of external osmolality on polyamine metabolism in HeLa cells. Biochim Biophys Acta. 1975 Dec 5;411(2):263–281. doi: 10.1016/0304-4165(75)90306-2. [DOI] [PubMed] [Google Scholar]
  21. Murakami Y., Fujita K., Kameji T., Hayashi S. Accumulation of ornithine decarboxylase-antizyme complex in HMOA cells. Biochem J. 1985 Feb 1;225(3):689–697. doi: 10.1042/bj2250689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Murakami Y., Matsufuji S., Miyazaki Y., Hayashi S. Forced expression of antizyme abolishes ornithine decarboxylase activity, suppresses cellular levels of polyamines and inhibits cell growth. Biochem J. 1994 Nov 15;304(Pt 1):183–187. doi: 10.1042/bj3040183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. 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]
  24. Perry J. W., Oka T. Regulation of ornithine decarboxylase in cultured mouse mammary gland by the osmolarity in the cellular environment. Biochim Biophys Acta. 1980 Apr 17;629(1):24–35. doi: 10.1016/0304-4165(80)90261-5. [DOI] [PubMed] [Google Scholar]
  25. Poulin R., Coward J. K., Lakanen J. R., Pegg A. E. Enhancement of the spermidine uptake system and lethal effects of spermidine overaccumulation in ornithine decarboxylase-overproducing L1210 cells under hyposmotic stress. J Biol Chem. 1993 Mar 5;268(7):4690–4698. [PubMed] [Google Scholar]
  26. Poulin R., Pegg A. E. Regulation of ornithine decarboxylase expression by anisosmotic shock in alpha-difluoromethylornithine-resistant L1210 cells. J Biol Chem. 1990 Mar 5;265(7):4025–4032. [PubMed] [Google Scholar]
  27. Poulin R., Wechter R. S., Pegg A. E. An early enlargement of the putrescine pool is required for growth in L1210 mouse leukemia cells under hypoosmotic stress. J Biol Chem. 1991 Apr 5;266(10):6142–6151. [PubMed] [Google Scholar]
  28. Rom E., Kahana C. Polyamines regulate the expression of ornithine decarboxylase antizyme in vitro by inducing ribosomal frame-shifting. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3959–3963. doi: 10.1073/pnas.91.9.3959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Suzuki T., He Y., Kashiwagi K., Murakami Y., Hayashi S., Igarashi K. Antizyme protects against abnormal accumulation and toxicity of polyamines in ornithine decarboxylase-overproducing cells. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8930–8934. doi: 10.1073/pnas.91.19.8930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tohyama Y., Kameji T., Hayashi S. Mechanisms of dramatic fluctuations of ornithine decarboxylase activity upon tonicity changes in primary cultured rat hepatocytes. Eur J Biochem. 1991 Dec 18;202(3):1327–1331. doi: 10.1111/j.1432-1033.1991.tb16507.x. [DOI] [PubMed] [Google Scholar]
  31. Tokunaga F., Goto T., Koide T., Murakami Y., Hayashi S., Tamura T., Tanaka K., Ichihara A. ATP- and antizyme-dependent endoproteolysis of ornithine decarboxylase to oligopeptides by the 26 S proteasome. J Biol Chem. 1994 Jul 1;269(26):17382–17385. [PubMed] [Google Scholar]
  32. Yang D., Hayashi H., Takii T., Mizutani Y., Inukai Y., Onozaki K. Interleukin-1-induced growth inhibition of human melanoma cells. Interleukin-1-induced antizyme expression is responsible for ornithine decarboxylase activity down-regulation. J Biol Chem. 1997 Feb 7;272(6):3376–3383. doi: 10.1074/jbc.272.6.3376. [DOI] [PubMed] [Google Scholar]
  33. van de Lest C. H., Veerkamp J. H., van Kuppevelt T. H. ImageCalc: a Microsoft Windows application for quantitative image analysis and comparison. Biotechniques. 1995 Jun;18(6):1050–1055. [PubMed] [Google Scholar]

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