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Nonsense-mediated mRNA decay occurs during eIF4F-dependent translation in human cells

Nature Structural & Molecular Biology volume 20, pages 702–709 (2013)Cite this article

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Abstract

The nonsense-mediated mRNA decay (NMD) pathway degrades mRNAs undergoing premature termination of translation. It has been argued that in human cells, NMD is restricted to a pioneer round of translation initiated on mRNAs associated with the cap-binding complex (CBC) and that the exchange of the CBC for the eIF4F translation initiation complex renders mRNAs immune to NMD. Here, we demonstrate that human mRNAs undergoing eIF4F-dependent translation are not immune to NMD. First, prolonged translation inhibition does not render an NMD substrate resistant to NMD, despite allowing exchange of CBC for eIF4F. Second, eIF4F inhibitors stabilize NMD substrates undergoing cap-dependent translation. Third, the eIF4E-associated pool of an NMD substrate degrades as rapidly as the overall pool of the mRNA. Thus, eIF4F-dependent translation supports NMD in human cells, allowing for the possibility that NMD could be activated upon cellular cues on already translating mRNAs.

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Figure 1: β39 mRNA remains competent for NMD after release from prolonged translation inhibition by cycloheximide or puromycin.
Figure 2: The degradation of β39 mRNA after release from puromycin-mediated translation inhibition is sensitive to 4EGI-1.
Figure 3: The eIF4F inhibitors 4EGI-1 and 4E-BP1 stabilize β39 mRNA but not β39 mRNA translated from an internal ribosome entry site.
Figure 4: 4EGI-1 and 4E-BP1 stabilize GPx1-46 and βWT-SMG5 NMD substrates.
Figure 5: The eIF4E-associated pool of β39 mRNA diminishes at the same rate as the overall β39 mRNA pool.
Figure 6: NMD can occur on mRNAs undergoing eIF4F-initiated translation in human cells.

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Acknowledgements

We thank P. Sarnow (Stanford University; pGL3-FLuc-IRES-Rluc), G. Wagner (Harvard Medical School; 4EGI-1) and E. Izaurralde (Max Planck Institute for Developmental Biology; antibody to CBP80) for valuable reagents. We thank R. Lardelli for critical comments on the manuscript and members of J.L.-A.'s laboratory for helpful discussions. This work was supported by a grant from the US National Science Foundation (MCB-0946464) to J.L.-A. and a postdoctoral fellowship from the Fondation pour la Recherche Médicale en France to S.D.

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  1. Division of Biological Sciences, University of California San Diego, La Jolla, California, USA

    Sébastien Durand & Jens Lykke-Andersen

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S.D. and J.L.-A. conceived of and designed the study. S.D. performed the experiments. Both authors analyzed the data and wrote the paper.

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Correspondence to Jens Lykke-Andersen.

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Durand, S., Lykke-Andersen, J. Nonsense-mediated mRNA decay occurs during eIF4F-dependent translation in human cells. Nat Struct Mol Biol 20, 702–709 (2013). https://doi.org/10.1038/nsmb.2575

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