Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2001 Sep 1;358(Pt 2):343–348. doi: 10.1042/0264-6021:3580343

Concurrent overexpression of ornithine decarboxylase and spermidine/spermine N(1)-acetyltransferase further accelerates the catabolism of hepatic polyamines in transgenic mice.

S Suppola 1, S Heikkinen 1, J J Parkkinen 1, M Uusi-Oukari 1, V P Korhonen 1, T Keinänen 1, L Alhonen 1, J Jänne 1
PMCID: PMC1222066  PMID: 11513732

Abstract

We have generated a hybrid transgenic mouse line overexpressing both ornithine decarboxylase (ODC) and spermidine/spermine N(1)-acetyltransferase (SSAT) under the control of the mouse metallothionein (MT) I promoter. In comparison with singly transgenic animals overexpressing SSAT, the doubly transgenic mice unexpectedly displayed much more striking signs of activated polyamine catabolism, as exemplified by a massive putrescine accumulation and an extreme reduction of hepatic spermidine and spermine pools. Interestingly, the profound depletion of the higher polyamines in the hybrid animals occurred in the presence of strikingly high ODC activity and tremendous putrescine accumulation. Polyamine catabolism in the doubly transgenic mice could be enhanced further by administration of zinc or the polyamine analogue N(1),N(11)-diethylnorspermine. In tracer experiments with [(14)C]spermidine we found that, in comparison with syngenic animals, both MT-ODC and MT-SSAT mice possessed an enhanced efflux mechanism for hepatic spermidine. In the MT-ODC animals this mechanism apparently operated in the absence of measurable SSAT activity. In the hybrid animals, spermidine efflux was stimulated further in comparison with the singly transgenic animals. In spite of a dramatic accumulation of putrescine and a profound reduction of the spermidine and spermine pools, only marginal changes were seen in the level of ODC antizyme. Even though the hybrid animals showed no liver or other organ-specific overt toxicity, except an early and permanent loss of hair, their life span was greatly reduced. These results can be understood from the perspective that catabolism is the overriding regulatory mechanism in the metabolism of the polyamines and that, even under conditions of severe depletion of spermidine and spermine, extremely high tissue pools of putrescine are not driven further to replenish the pools of the higher polyamines.

Full Text

The Full Text of this article is available as a PDF (111.6 KB).

Selected References

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

  1. Alhonen L., Halmekytö M., Kosma V. M., Wahlfors J., Kauppinen R., Jänne J. Life-long over-expression of ornithine decarboxylase (ODC) gene in transgenic mice does not lead to generally enhanced tumorigenesis or neuronal degeneration. Int J Cancer. 1995 Nov 3;63(3):402–404. doi: 10.1002/ijc.2910630317. [DOI] [PubMed] [Google Scholar]
  2. Alhonen L., Heikkinen S., Sinervirta R., Halmekytö M., Alakuijala P., Jänne J. Transgenic mice expressing the human ornithine decarboxylase gene under the control of mouse metallothionein I promoter. Biochem J. 1996 Mar 1;314(Pt 2):405–408. doi: 10.1042/bj3140405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alhonen L., Karppinen A., Uusi-Oukari M., Vujcic S., Korhonen V. P., Halmekytö M., Kramer D. L., Hines R., Jänne J., Porter C. W. Correlation of polyamine and growth responses to N1,N11-diethylnorspermine in primary fetal fibroblasts derived from transgenic mice overexpressing spermidine/spermine N1-acetyltransferase. J Biol Chem. 1998 Jan 23;273(4):1964–1969. doi: 10.1074/jbc.273.4.1964. [DOI] [PubMed] [Google Scholar]
  4. Alhonen L., Parkkinen J. J., Keinanen T., Sinervirta R., Herzig K. H., Jänne J. Activation of polyamine catabolism in transgenic rats induces acute pancreatitis. Proc Natl Acad Sci U S A. 2000 Jul 18;97(15):8290–8295. doi: 10.1073/pnas.140122097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Alhonen L., Pietilä M., Halmekytö M., Kramer D. L., Jänne J., Porter C. W. Transgenic mice with activated polyamine catabolism due to overexpression of spermidine/spermine N1-acetyltransferase show enhanced sensitivity to the polyamine analog, N1, N11-diethylnorspermine. Mol Pharmacol. 1999 Apr;55(4):693–698. [PubMed] [Google Scholar]
  6. Bernacki R. J., Oberman E. J., Seweryniak K. E., Atwood A., Bergeron R. J., Porter C. W. Preclinical antitumor efficacy of the polyamine analogue N1, N11-diethylnorspermine administered by multiple injection or continuous infusion. Clin Cancer Res. 1995 Aug;1(8):847–857. [PubMed] [Google Scholar]
  7. Casero R. A., Jr, Pegg A. E. Spermidine/spermine N1-acetyltransferase--the turning point in polyamine metabolism. FASEB J. 1993 May;7(8):653–661. [PubMed] [Google Scholar]
  8. Hakovirta H., Keiski A., Toppari J., Halmekytö M., Alhonen L., Jänne J., Parvinen M. Polyamines and regulation of spermatogenesis: selective stimulation of late spermatogonia in transgenic mice overexpressing the human ornithine decarboxylase gene. Mol Endocrinol. 1993 Nov;7(11):1430–1436. doi: 10.1210/mend.7.11.8114757. [DOI] [PubMed] [Google Scholar]
  9. Halmekytö M., Alhonen L., Alakuijala L., Jänne J. Transgenic mice over-producing putrescine in their tissues do not convert the diamine into higher polyamines. Biochem J. 1993 Apr 15;291(Pt 2):505–508. doi: 10.1042/bj2910505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Halmekytö M., Alhonen L., Wahlfors J., Sinervirta R., Eloranta T., Jänne J. Characterization of a transgenic mouse line over-expressing the human ornithine decarboxylase gene. Biochem J. 1991 Sep 15;278(Pt 3):895–898. doi: 10.1042/bj2780895. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Halmekytö M., Hyttinen J. M., Sinervirta R., Utriainen M., Myöhänen S., Voipio H. M., Wahlfors J., Syrjänen S., Syrjänen K., Alhonen L. Transgenic mice aberrantly expressing human ornithine decarboxylase gene. J Biol Chem. 1991 Oct 15;266(29):19746–19751. [PubMed] [Google Scholar]
  12. Halmekytö M., Syrjänen K., Jänne J., Alhonen L. Enhanced papilloma formation in response to skin tumor promotion in transgenic mice overexpressing the human ornithine decarboxylase gene. Biochem Biophys Res Commun. 1992 Aug 31;187(1):493–497. doi: 10.1016/s0006-291x(05)81521-9. [DOI] [PubMed] [Google Scholar]
  13. Halonen T., Sivenius J., Miettinen R., Halmekytö M., Kauppinen R., Sinervirta R., Alakuijala L., Alhonen L., MacDonald E., Jänne J. Elevated seizure threshold and impaired spatial learning in transgenic mice with putrescine overproduction in the brain. Eur J Neurosci. 1993 Sep 1;5(9):1233–1239. doi: 10.1111/j.1460-9568.1993.tb00978.x. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Heljasvaara R., Veress I., Halmekytö M., Alhonen L., Jänne J., Laajala P., Pajunen A. Transgenic mice overexpressing ornithine and S-adenosylmethionine decarboxylases maintain a physiological polyamine homoeostasis in their tissues. Biochem J. 1997 Apr 15;323(Pt 2):457–462. doi: 10.1042/bj3230457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. 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]
  17. Hyvönen T., Keinänen T. A., Khomutov A. R., Khomutov R. M., Eloranta T. O. Monitoring of the uptake and metabolism of aminooxy analogues of polyamines in cultured cells by high-performance liquid chromatography. J Chromatogr. 1992 Feb 7;574(1):17–21. doi: 10.1016/0378-4347(92)80093-6. [DOI] [PubMed] [Google Scholar]
  18. Jänne J., Williams-Ashman H. G. Dissociation of putrescine-activated decarboxylation of S-adenosyl-L-methionine from the enzymic synthesis of spermidine and spermine by purified prostatic enzyme preparations. Biochem Biophys Res Commun. 1971 Jan 22;42(2):222–229. [PubMed] [Google Scholar]
  19. Jänne J., Williams-Ashman H. G. On the purification of L-ornithine decarboxylase from rat prostate and effects of thiol compounds on the enzyme. J Biol Chem. 1971 Mar 25;246(6):1725–1732. [PubMed] [Google Scholar]
  20. Kaasinen K., Koistinaho J., Alhonen L., Jänne J. Overexpression of spermidine/spermine N-acetyltransferase in transgenic mice protects the animals from kainate-induced toxicity. Eur J Neurosci. 2000 Feb;12(2):540–548. doi: 10.1046/j.1460-9568.2000.00940.x. [DOI] [PubMed] [Google Scholar]
  21. Kankare K., Uusi-Oukari M., Jänne O. A. Structure, organization and expression of the mouse ornithine decarboxylase antizyme gene. Biochem J. 1997 Jun 15;324(Pt 3):807–813. doi: 10.1042/bj3240807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lukkarinen J. A., Kauppinen R. A., Gröhn O. H., Oja J. M., Sinervirta R., Järvinen A., Alhonen L. I., Jänne J. Neuroprotective role of ornithine decarboxylase activation in transient focal cerebral ischaemia: a study using ornithine decarboxylase-overexpressing transgenic rats. Eur J Neurosci. 1998 Jun;10(6):2046–2055. doi: 10.1046/j.1460-9568.1998.00216.x. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. 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]
  25. Nilsson J., Koskiniemi S., Persson K., Grahn B., Holm I. Polyamines regulate both transcription and translation of the gene encoding ornithine decarboxylase antizyme in mouse. Eur J Biochem. 1997 Dec 1;250(2):223–231. doi: 10.1111/j.1432-1033.1997.0223a.x. [DOI] [PubMed] [Google Scholar]
  26. Pietilä M., Alhonen L., Halmekytö M., Kanter P., Jänne J., Porter C. W. Activation of polyamine catabolism profoundly alters tissue polyamine pools and affects hair growth and female fertility in transgenic mice overexpressing spermidine/spermine N1-acetyltransferase. J Biol Chem. 1997 Jul 25;272(30):18746–18751. doi: 10.1074/jbc.272.30.18746. [DOI] [PubMed] [Google Scholar]
  27. Rehse K., Puchert E., Leissring S. Antiaggregatorische und anticoagulante Eigenschaften von Oligoaminen, 12. Mitt. Alkyl- und Arylalkylderivate von Putrescin, Spermidin und Spermin. Arch Pharm (Weinheim) 1990 May;323(5):287–294. doi: 10.1002/ardp.19903230507. [DOI] [PubMed] [Google Scholar]
  28. Suppola S., Pietilä M., Parkkinen J. J., Korhonen V. P., Alhonen L., Halmekytö M., Porter C. W., Jänne J. Overexpression of spermidine/spermine N1-acetyltransferase under the control of mouse metallothionein I promoter in transgenic mice: evidence for a striking post-transcriptional regulation of transgene expression by a polyamine analogue. Biochem J. 1999 Mar 1;338(Pt 2):311–316. [PMC free article] [PubMed] [Google Scholar]
  29. Vujcic S., Halmekyto M., Diegelman P., Gan G., Kramer D. L., Janne J., Porter C. W. Effects of conditional overexpression of spermidine/spermine N1-acetyltransferase on polyamine pool dynamics, cell growth, and sensitivity to polyamine analogs. J Biol Chem. 2000 Dec 8;275(49):38319–38328. doi: 10.1074/jbc.M003270200. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES