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Polyamines pp 279-292 | Cite as

Posttranscriptional Regulation of Ornithine Decarboxylase

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

Abstract

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 

Notes

Acknowledgements

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).

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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

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