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A novel mode of RBD-protein recognition in the Y14–Mago complex

Nature Structural & Molecular Biology volume 10, pages 433–439 (2003)Cite this article

Abstract

Y14 and Mago are conserved eukaryotic proteins that associate with spliced mRNAs in the nucleus and remain associated at exon junctions during and after nuclear export. In the cytoplasm, Y14 is involved in mRNA quality control via the nonsense-mediated mRNA decay (NMD) pathway and, together with Mago, is involved in localization of osk (oskar) mRNA. We have determined the crystal structure of the complex between Drosophila melanogaster Y14 and Mago at a resolution of 2.5 Å. The structure reveals an atypical mode of protein–protein recognition mediated by an RNA-binding domain (RBD). Instead of binding RNA, the RBD of Y14 engages its RNP1 and RNP2 motifs to bind Mago. Using structure-guided mutagenesis, we show that Mago is also a component of the NMD pathway, and that its association with Y14 is essential for function. Heterodimerization creates a single structural platform that interacts with the NMD machinery via phylogenetically conserved residues.

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Figure 1: Structure of the Y14–Mago heterodimer.
Figure 2: Structure-based amino acid sequence alignments of Mago and Y14.
Figure 3: Main heterodimerization interface between Y14 and Mago.
Figure 4: Activity of human Y14 and Mago mutants in NMD.
Figure 5: Conserved molecular surfaces of the Y14–Mago heterodimer are involved in NMD.
Figure 6: RNA-binding properties of human Y14 and Mago.

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Acknowledgements

We are grateful to beamline scientists at DESY BW7A (Hamburg), Swiss Light Source X06SA (Zurich), Elettra (Trieste) and ESRF ID14-4 and ID14-1 (Grenoble) for assistance during data collection. We thank in particular M. Polentarutti and K. Djinovic (Elettra) for help with xenon derivatization. We thank N. Gehring, A. Kulozik and M. Hentze for communicating results on the NMD reporter assay before publication, and for the gift of the reporter constructs and λN-peptide–specific antibodies. We also thank I. Mattaj, P. Brick and A. Ladurner for critical reading of the manuscript. S.F. was supported by a Marie Curie Fellowship.

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  1. European Molecular Biology Laboratory, Meyerhofstrasse 1, Heidelberg, D-69117, Germany

    Sébastien Fribourg, David Gatfield, Elisa Izaurralde & Elena Conti

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Correspondence to Elena Conti.

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Fribourg, S., Gatfield, D., Izaurralde, E. et al. A novel mode of RBD-protein recognition in the Y14–Mago complex. Nat Struct Mol Biol 10, 433–439 (2003). https://doi.org/10.1038/nsb926

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