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Nucleic Acids Research 2006 34(8):2355-2363; doi:10.1093/nar/gkl277
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Published online 5 May 2006

© The Author 2006. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions@oxfordjournals.org

Article

The uORF-containing thrombopoietin mRNA escapes nonsense-mediated decay (NMD)

Clemens Stockklausner1,2, Stephen Breit1,2, Gabriele Neu-Yilik1,2, Nicole Echner1,2, Matthias W. Hentze2,3, Andreas E. Kulozik1,2 and Niels H. Gehring1,2

1 Department of Pediatric Oncology, Hematology and Immunology, University of Heidelberg 69120 Heidelberg, Germany 2 Molecular Medicine Partnership Unit, EMBL/University of Heidelberg 69120 Heidelberg, Germany 3 European Molecular Biology Laboratory; Gene Expression Unit 69117 Heidelberg, Germany

To whom correspondence should be addressed at Im Neuenheimer Feld 156, 69120 Heidelberg, Germany. Tel: +49 6221 56 4581; Fax: +49 6221 56 4580; Email: Niels.Gehring{at}med.uni-heidelberg.de

Received January 23, 2006. Revised February 8, 2006. Accepted April 4, 2006.

Platelet production is induced by the cytokine thrombopoietin (TPO). It is physiologically critical that TPO expression is tightly regulated, because lack of TPO causes life-threatening thrombocytopenia while an excess of TPO results in thrombocytosis. The plasma concentration of TPO is controlled by a negative feedback loop involving receptor-mediated uptake of TPO by platelets. Furthermore, TPO biosynthesis is limited by upstream open reading frames (uORFs) that curtail the translation of the TPO mRNA. uORFs are suggested to activate RNA degradation by nonsense-mediated decay (NMD) in a number of physiological transcripts. Here, we determine whether NMD affects TPO expression. We show that reporter mRNAs bearing the seventh TPO uORF escape NMD. Importantly, endogenously expressed TPO mRNA from HuH7 cells is unaffected by abrogation of NMD by RNAi. Thus, regulation of TPO expression is independent of NMD, implying that mRNAs bearing uORFs cannot generally be considered to represent NMD targets.


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