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eIF4E-bound mRNPs are substrates for nonsense-mediated mRNA decay in mammalian cells

Nature Structural & Molecular Biology volume 20, pages 710–717 (2013)Cite this article

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Abstract

Eukaryotic mRNAs with premature translation termination codons (PTCs) are recognized and degraded through a process termed nonsense-mediated mRNA decay (NMD). The evolutionary conservation of the core NMD factors UPF1, UPF2 and UPF3 implies a similar basic mechanism of PTC recognition in all eukaryotes. However, while PTC-containing mRNAs in yeast seem to be available to NMD at each round of translation, mammalian NMD has been reported to be restricted to cap-binding complex (CBC)–bound mRNAs during the pioneer round of translation. Here, we compared decay kinetics of two NMD reporter genes in mRNA fractions bound to either CBC or the eukaryotic initiation factor 4E (eIF4E) in human cells and demonstrate that NMD destabilizes eIF4E-bound transcripts as efficiently as those associated with CBC. These results corroborate an emerging unified model for NMD substrate recognition, according to which NMD can ensue at every aberrant translation termination event.

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Figure 1: Immunopurification of CBC- and eIF4E-mRNPs.
Figure 2: CBP80- and eIF4E-associated PTC-containing TCR-β mRNAs are NMD targets.
Figure 3: CBP80- and eIF4E-associated PTC+ miniμ reporter mRNA is rapidly degraded by NMD when the poly(A) tail is distant from the PTC.
Figure 4: NMD targets both CBC- and eIF4E-bound mRNAs: a graphical summary.

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Acknowledgements

We thank E. Izaurralde (Max Planck Institute), N. Sonenberg (McGill University) and Y.-K. Kim (Korea University) for antibodies, and J. Lykke-Andersen and A. Jacobson for valuable comments on the manuscript. This work was supported by grants of the European Research Council (StG 207419; O.M.), the Swiss National Science Foundation (31003A-127614 and 31003A-143717; O.M.) and the canton of Bern.

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  1. Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland

    Simone C Rufener & Oliver Mühlemann

  2. Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland

    Simone C Rufener

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S.C.R. and O.M. conceived of and designed the experiments. S.C.R. performed the experiments. O.M. and S.C.R wrote the paper. O.M. provided resources.

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Correspondence to Oliver Mühlemann.

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Rufener, S., Mühlemann, O. eIF4E-bound mRNPs are substrates for nonsense-mediated mRNA decay in mammalian cells. Nat Struct Mol Biol 20, 710–717 (2013). https://doi.org/10.1038/nsmb.2576

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