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PYM binds the cytoplasmic exon-junction complex and ribosomes to enhance translation of spliced mRNAs

Nature Structural & Molecular Biology volume 14, pages 1173–1179 (2007)Cite this article

Abstract

Messenger RNAs produced by splicing are translated more efficiently than those produced from similar intronless precursor mRNAs (pre-mRNAs). The exon-junction complex (EJC) probably mediates this enhancement; however, the specific link between the EJC and the translation machinery has not been identified. The EJC proteins Y14 and magoh remain bound to spliced mRNAs after their export from the nucleus to the cytoplasm and are removed only when these mRNAs are translated. Here we show that PYM, a 29-kDa protein that binds the Y14–magoh complex in the cytoplasm, also binds, via a separate domain, to the small (40S) ribosomal subunit and the 48S preinitiation complex. Furthermore, PYM knockdown reduces the translation efficiency of a reporter protein produced from intron-containing, but not intronless, pre-mRNA. We suggest that PYM functions as a bridge between EJC-bearing spliced mRNAs and the translation machinery to enhance translation of the mRNAs.

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Figure 1: PYM interacts selectively with the Y14–magoh complex.
Figure 2: PYM associates with the 40S ribosomal subunit and translation factors.
Figure 3: The C terminus of PYM is required for association with the 48S preinitiation complex.
Figure 4: Knockdown of PYM results in reduced expression from an intron-containing reporter.
Figure 5: Model of the assembly pathway and dynamics of the EJC and the role of PYM in the interaction of the EJC with the translation machinery.

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Acknowledgements

We thank members of our laboratory, especially D. Battle and J. Yong, for stimulating discussions and comments on this manuscript; H. Sheng and R. Olszewski for technical assistance; S. Grill for secretarial assistance; I. Mattaj (European Molecular Biology Laboratory) for the CBP20 and CBP80 antibodies; G. Van Duyne (University of Pennsylvania) for the GST-TEV expression vector; and the New York University Protein Analysis Facility for protein microsequencing. G.D. is an Investigator of the Howard Hughes Medical Institute.

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  1. Department of Biochemistry and Biophysics, Howard Hughes Medical Institute, University of Pennsylvania School of Medicine, Philadelphia, 19104-6148, Pennsylvania, USA

    Michael D Diem, Chia C Chan, Ihab Younis & Gideon Dreyfuss

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  1. Michael D Diem
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Correspondence to Gideon Dreyfuss.

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Diem, M., Chan, C., Younis, I. et al. PYM binds the cytoplasmic exon-junction complex and ribosomes to enhance translation of spliced mRNAs. Nat Struct Mol Biol 14, 1173–1179 (2007). https://doi.org/10.1038/nsmb1321

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