Polyamines pp 279-292 | Cite as

Posttranscriptional Regulation of Ornithine Decarboxylase

  • Shannon L. Nowotarski
  • Sofia Origanti
  • Lisa M. ShantzEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 720)


Activity of the polyamine biosynthetic enzyme ornithine decarboxylase (ODC) and intracellular levels of ODC protein are controlled very tightly. Numerous studies have described ODC regulation at the levels of transcription, translation, and protein degradation in normal cells and dysregulation of these processes in response to oncogenic stimuli. Although posttranscriptional regulation of ODC has been well documented, the RNA binding proteins (RBPs) that interact with ODC mRNA and control synthesis of the ODC protein have not been defined. Using Ras-transformed rat intestinal epithelial cells (Ras12V cells) as a model, we have begun identifying the RBPs that associate with the ODC transcript. Binding of RBPs could potentially regulate ODC synthesis by either changing mRNA stability or rate of mRNA translation. Techniques for measuring RBP binding and translation initiation are described here. Targeting control of ODC translation or mRNA decay could be a valuable method of limiting polyamine accumulation and subsequent tumor development in a variety of cancers.

Key words

Ornithine decarboxylase Polyamines RNA stability Protein synthesis Translational regulation Polysome profiles mRNP assay AU-rich region HuR 



The authors would like to thank Suzanne Sass-Kuhn for excellent technical assistance. This work was supported by R01 CA82768 and R03 CA142051 (to LMS).


  1. 1.
    Pegg AE (2006) Regulation of ornithine decarboxylase. J Biol Chem 281:4529–14532PubMedCrossRefGoogle Scholar
  2. 2.
    Zhao B, Butler AP (2001) Core promoter involvement in the induction of rat ornithine decarboxylase by phorbol esters. Mol Carcinog 32:92–99PubMedCrossRefGoogle Scholar
  3. 3.
    Li R, Abrahamsen MS, Johnson RR, Morris DR (1994) Complex interactions at a GC-rich domain regulate cell type-dependent activity of the ornithine decarboxylase promoter. J Biol Chem 269:7941–7949PubMedGoogle Scholar
  4. 4.
    Wallon UM, Persson L, Heby O (1995) Regulation of ornithine decarboyxlase during cell growth. Changes in the stability and translatability of the mRNA, and in the turnover of the protein. Mol Cell Biochem 146:39–44PubMedCrossRefGoogle Scholar
  5. 5.
    Shantz LM (2004) Transcriptional and translational control of ornithine decarboxylase during Ras transformation. Biochem J 377:257–264PubMedCrossRefGoogle Scholar
  6. 6.
    Shantz LM, Pegg AE (1999) Translational regulation of ornithine decarboxylase and other enzymes of the polyamine pathway. Int J Biochem Cell Biol 31:107–122PubMedCrossRefGoogle Scholar
  7. 7.
    Huang Y, Pledgie A, Casero RA Jr, Davidson NE (2005) Molecular mechanisms of polyamine analogs in cancer cells. Anticancer Drugs 16:229–241PubMedCrossRefGoogle Scholar
  8. 8.
    Gerner EW, Meyskens FL Jr (2004) Polyamines and cancer: old molecules, new understanding. Nat Rev Cancer 4:781–792PubMedCrossRefGoogle Scholar
  9. 9.
    O’Brien TG, Megosh LC, Gilliard G, Peralta Soler A (1997) Ornithine decarboxylase overexpression is a sufficent condition for tumor promotion in mouse skin. Cancer Res 57:2630–2637PubMedGoogle Scholar
  10. 10.
    O’Brien TG (1976) The induction of ornithine decarboxylase is an early, possibly obligatory event in mouse skin carcinogenesis. Cancer Res 36:2644–2653PubMedGoogle Scholar
  11. 11.
    Origanti S, Shantz LM (2007) Ras transformation of RIE-1 cells activates cap-independent translation of ornithine decarboxylase: regulation by the Raf/MEK/ERK and phosphatidylinositol 3-kinase pathways. Cancer Res 67:4834–4842PubMedCrossRefGoogle Scholar
  12. 12.
    Shantz LM, Pegg AE (1994) Overproduction of ornithine decarboxylase caused by relief of translational repression is associated with neoplastic transformation. Cancer Res 54:2313–2316PubMedGoogle Scholar
  13. 13.
    Rousseau D, Kaspar R, Rosenwald I, Gehrke L, Sonenberg N (1996) Translation initiation of ornithine decarboxylase and nucleocytoplasmic transport of cyclin D1 mRNA are increased in cells overexpressing eukaryotic initiation factor 4E. Proc Natl Acad Sci USA 93:1065–1070PubMedCrossRefGoogle Scholar
  14. 14.
    Grens A, Scheffler IE (1990) The 5’- and 3’-untranslated regions of ornithine decarboxylase mRNA affect the translational efficiency. J Biol Chem 265:11810–11816PubMedGoogle Scholar
  15. 15.
    Lorenzini EC, Scheffler IE (1997) Co-operation of the 5’ and 3’ untranslated regions of ornithine decarboxylase mRNA and inhibitory role of its 3’ untranslated region in regulating the translational efficiency of hybrid RNA species via cellular factors. Biochem J 326:361–367PubMedGoogle Scholar
  16. 16.
    Lövkvist-Wallstrom E, Stjernborg-Ulvsbäck L, Scheffler IE, Persson L (1995) Regulation of mammalian ornithine decarboxylase. Studies on the induction of the enzyme by hypotonic stress. Eur J Biochem 231:40–44PubMedCrossRefGoogle Scholar
  17. 17.
    Zou T, Mazan-Mamczarz K, Rao JN, Liu L, Marasa BS, Zhang AH, Xiao L, Pullmann R, Gorospe M, Wang JY (2006) Polyamine depletion increases cytoplasmic levels of RNA-binding protein HuR leading to stabilization of nucleophosmin and p53 mRNAs. J Biol Chem 281:19387–19394PubMedCrossRefGoogle Scholar
  18. 18.
    Xiao L, Rao JN, Zou T, Liu L, Marasa BS, Chen J, Turner DJ, Zhou H, Gorospe M, Wang JY (2007) Polyamines regulate the stability of activating transcription factor-2 mRNA through RNA-binding protein HuR in intestinal epithelial cells. Mol Biol Cell 18:4579–4590PubMedCrossRefGoogle Scholar
  19. 19.
    Garneau NL, Wilusz J, Wilusz CJ (2007) The highways and byways of mRNA decay. Nat Rev Mol Cell Biol 8:113–126PubMedCrossRefGoogle Scholar
  20. 20.
    Barreau C, Paillard L, Osborne HB (2005) AU-rich elements and associated factors: are there unifying principles? Nucleic Acids Res 33:7138–7150PubMedCrossRefGoogle Scholar
  21. 21.
    Hau HH, Walsh RJ, Ogilvie RL, Williams DA, Reilly CS, Bohjanen PR (2007) Tristetraprolin recruits functional mRNA decay complexes to ARE sequences. J Cell Biochem 100:1477–1492PubMedCrossRefGoogle Scholar
  22. 22.
    Parker R, Sheth U (2007) P bodies and the control of mRNA translation and degradation. Mol Cell 25:635–646PubMedCrossRefGoogle Scholar
  23. 23.
    Eulalio A, Behm-Ansmant I, Izaurralde E (2007) P bodies: at the crossroads of post-transcriptional pathways. Nat Rev Mol Cell Biol 8:9–22PubMedCrossRefGoogle Scholar
  24. 24.
    Johannes G, Carter MS, Eisen MB, Brown PO, Sarnow P (1999) Identification of eukaryotic mRNAs that are translated at reduced cap binding complex eIF4F concentrations using a cDNA microarray. Proc Natl Acad Sci USA 96:13118–13123PubMedCrossRefGoogle Scholar
  25. 25.
    Bradford M (1976) A rapid sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  26. 26.
    Lu L, Stanley BA, Pegg AE (1991) Identification of residues in ornithine decarboxylase essential for enzymatic activity and for rapid protein turnover. Biochem J 277:671–675PubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Shannon L. Nowotarski
    • 1
  • Sofia Origanti
    • 1
  • Lisa M. Shantz
    • 1
    Email author
  1. 1.Department of Cellular and Molecular Physiology, Milton S. Hershey Medical CenterPennsylvania State University College of MedicineHersheyUSA

Personalised recommendations