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A direct interaction between DCP1 and XRN1 couples mRNA decapping to 5′ exonucleolytic degradation

Abstract

The removal of the mRNA 5′ cap structure by the decapping enzyme DCP2 leads to rapid 5′→3′ mRNA degradation by XRN1, suggesting that the two processes are coordinated, but the coupling mechanism is unknown. DCP2 associates with the decapping activators EDC4 and DCP1. Here we show that XRN1 directly interacts with EDC4 and DCP1 in human and Drosophila melanogaster cells, respectively. In D. melanogaster cells, this interaction is mediated by the DCP1 EVH1 domain and a DCP1-binding motif (DBM) in the XRN1 C-terminal region. The NMR structure of the DCP1 EVH1 domain bound to the DBM reveals that the peptide docks at a conserved aromatic cleft, which is used by EVH1 domains to recognize proline-rich ligands. Our findings reveal a role for XRN1 in decapping and provide a molecular basis for the coupling of decapping to 5′→3′ mRNA degradation.

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Figure 1: XRN1 interacts with DCP1 in D. melanogaster cells.
Figure 2: The structure of the DCP1 EVH1 domain bound to the XRN1 DBM motif.
Figure 3: XRN1 interacts with EDC4 in human cells.
Figure 4: Mutagenesis of the D. melanogaster DCP1-XRN1 interaction interface.
Figure 5: XRN1 overexpression inhibits mRNA decapping.
Figure 6: The DCP1-XRN1 interaction is required for efficient decapping in vivo.

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Acknowledgements

We are grateful to S.F. Newbury (Brighton and Sussex Medical School, University of Sussex, Brighton, UK) for the kind gift of D. melanogaster XRN1 complementary DNA (cDNA) and XRN1 antibodies. This study was supported by the Max Planck Society and grants from the Deutsche Forschungsgemeinschaft (DFG, FOR855 to E.I. and the Gottfried Wilhelm Leibniz Program to E.I.).

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Contributions

E.I. conceived the project. J.E.B. and E.H. carried out the immunoprecipitations and functional assays in S2 cells. J.E.B. and A.B. purified the complex. C.-T.C. carried out immunoprecipitations in human cells. G.H. was involved in an earlier phase of the project, cloned XRN1 and DCP1 fragments and performed preliminary immunoprecipitations and functional assays in S2 cells. V.T. collected and processed the NMR data. V.T. and M.C. solved the structure and built the model. J.E.B., V.T., A.B., M.C., O.W. and E.I. analyzed the structure and wrote the manuscript.

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Correspondence to Vincent Truffault or Elisa Izaurralde.

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Braun, J., Truffault, V., Boland, A. et al. A direct interaction between DCP1 and XRN1 couples mRNA decapping to 5′ exonucleolytic degradation. Nat Struct Mol Biol 19, 1324–1331 (2012). https://doi.org/10.1038/nsmb.2413

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