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Analysis of Cotranslational Polyamine Sensing During Decoding of ODC Antizyme mRNA

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Book cover Polyamines

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1694))

Abstract

Polyamines are essential poly-cations with vital functions in all cellular systems. Their levels are controlled by intricate regulatory feedback mechanisms. Abnormally high levels of polyamines have been linked to cancer. A rate-limiting enzyme in the biosynthesis of polyamines in fungi and higher eukaryotes is ornithine-decarboxylase (ODC). Its levels are largely controlled posttranslationally via ubiquitin-independent degradation mediated by ODC antizyme (OAZ). The latter is a critical polyamine sensor in a feedback control mechanism that adjusts cellular polyamine levels. Here, we describe an approach employing quantitative western blot analyses that provides in vivo evidence for cotranslational polyamine-sensing by nascent OAZ in yeast. In addition, we describe an in vitro method to detect polyamine binding by antizyme.

The original version of this chapter was revised. A correction to this chapter can be found at https://doi.org/10.1007/978-1-4939-7398-9_41

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  • 06 September 2018

    Correction to: Chapter 26 in: Rubén Alcázar and Antonio F. Tiburcio (eds.), Polyamines: Methods and Protocols, Methods in Molecular Biology, vol. 1694, https://doi.org/10.1007/978-1-4939-7398-9_26

References

  1. Michael AJ (2016) Biosynthesis of polyamines and polyamine-containing molecules. Biochem J 473(15):2315–2329

    Article  CAS  PubMed  Google Scholar 

  2. Tabor CW, Tabor H (1984) Polyamines. Annu Rev Biochem 53:749–790

    Article  CAS  PubMed  Google Scholar 

  3. Thomas T, Thomas TJ (2001) Polyamines in cell growth and cell death: molecular mechanisms and therapeutic applications. Cell Mol Life Sci 58:244–258

    Article  CAS  PubMed  Google Scholar 

  4. Childs AC, Mehta DJ, Gerner EW (2003) Polyamine-dependent gene expression. Cell Mol Life Sci 60:1394–1406

    Article  CAS  PubMed  Google Scholar 

  5. Palanimurugan R, Kurian L, Hegde V, Hofmann K, Dohmen RJ (2014) Co-translational polyamine sensing by nascent ODC Antizyme. In: Ito K (ed) Regulatory nascent polypeptides. Springer, Tokyo, pp 203–222

    Google Scholar 

  6. Nakanishi S, Cleveland JL (2016) Targeting the polyamine-hypusine circuit for the prevention and treatment of cancer. Amino Acids 48:2353–2362

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Murray-Stewart TR, Woster PM, Casero RA Jr (2016) Targeting polyamine metabolism for cancer therapy and prevention. Biochem J 473:2937–2953

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Liu JH, Wang W, Wu H, Gong X, Moriguchi T (2015) Polyamines function in stress tolerance: from synthesis to regulation. Front Plant Sci 6:827

    PubMed  PubMed Central  Google Scholar 

  9. Tiburcio AF, Altabella T, Bitrian M, Alcazar R (2014) The roles of polyamines during the lifespan of plants: from development to stress. Planta 240:1–18

    Article  CAS  Google Scholar 

  10. Coffino P (2001) Regulation of cellular polyamines by antizyme. Nat Rev Mol Cell Biol 2:188–194

    Article  CAS  PubMed  Google Scholar 

  11. Illingworth C, Michael AJ (2012) Plant ornithine decarboxylase is not post-transcriptionally feedback regulated by polyamines but can interact with a cytosolic ribosomal protein S15 polypeptide. Amino Acids 42:519–527

    Article  CAS  PubMed  Google Scholar 

  12. Kahana C (2016) Protein degradation, the main hub in the regulation of cellular polyamines. Biochem J 473:4551–4558

    Article  CAS  PubMed  Google Scholar 

  13. Murakami Y, Matsufuji S, Kameji T, Hayashi S, Igarashi K, Tamura T, Tanaka K, Ichihara A (1992) Ornithine decarboxylase is degraded by the 26S proteasome without ubiquitination. Nature 360:597–599

    Article  CAS  PubMed  Google Scholar 

  14. Palanimurugan R, Scheel H, Hofmann K, Dohmen RJ (2004) Polyamines regulate their synthesis by inducing expression and blocking degradation of ODC antizyme. EMBO J 23:4857–4867

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Matsufuji S, Matsufuji T, Miyazaki Y, Murakami Y, Atkins JF, Gesteland RF, Hayashi S (1995) Autoregulatory frameshifting in decoding mammalian ornithine decarboxylase antizyme. Cell 80:51–60

    Article  CAS  PubMed  Google Scholar 

  16. Ivanov IP, Atkins JF (2007) Ribosomal frameshifting in decoding antizyme mRNAs from yeast and protists to humans: close to 300 cases reveal remarkable diversity despite underlying conservation. Nucleic Acids Res 35:1842–1858

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Kurian L, Palanimurugan R, Gödderz D, Dohmen RJ (2011) Polyamine sensing by nascent ornithine decarboxylase antizyme stimulates decoding of its mRNA. Nature 477:490–494

    Article  CAS  PubMed  Google Scholar 

  18. Heinemeyer W, Kleinschmidt JA, Saidowsky J, Escher C, Wolf DH (1991) Proteinase yscE, the yeast proteasome/multicatalytic-multifunctional proteinase: mutants unravel its function in stress induced proteolysis and uncover its necessity for cell survival. EMBO J 10:555–562

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  19. Gietz RD, Sugino A (1988) New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74:527–534

    Article  CAS  PubMed  Google Scholar 

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Author information

Authors and Affiliations

  1. Center for Cellular and Molecular Biology (CCMB), Hyderabad, 500007, India

    R. Palanimurugan

  2. Deallus Consulting Ltd, 1 Swan Lane, London, EC4R 3TN, UK

    Daniela Gödderz

  3. Laboratory for Developmental and Regenerative RNA biology, Center of Molecular Medicine Cologne (CMMC), University of Cologne, 50931, Cologne, Germany

    Leo Kurian

  4. Institute for Genetics, Biocenter, University of Cologne, 50674, Cologne, Germany

    R. J. Dohmen

Authors
  1. R. Palanimurugan
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  2. Daniela Gödderz
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  3. Leo Kurian
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  4. R. J. Dohmen
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Corresponding author

Correspondence to R. J. Dohmen .

Editor information

Editors and Affiliations

  1. Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Section of Plant Physiology, University of Barcelona, Spain

    Rubén Alcázar

  2. Department of Biology, Healthcare and Environment, Faculty of Pharmacy and Food Sciences, Section of Plant Physiology, University of Barcelona, Spain

    Antonio F. Tiburcio

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Palanimurugan, R., Gödderz, D., Kurian, L., Dohmen, R.J. (2018). Analysis of Cotranslational Polyamine Sensing During Decoding of ODC Antizyme mRNA. In: Alcázar, R., Tiburcio, A. (eds) Polyamines. Methods in Molecular Biology, vol 1694. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7398-9_26

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  • DOI: https://doi.org/10.1007/978-1-4939-7398-9_26

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7397-2

  • Online ISBN: 978-1-4939-7398-9

  • eBook Packages: Springer Protocols

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