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Gemin5-snRNA interaction reveals an RNA binding function for WD repeat domains

Nature Structural & Molecular Biology volume 16, pages 486–491 (2009)Cite this article

Abstract

Gemin5 binds specifically to the small nuclear RNA (snRNA)-defining small nuclear ribonucleoprotein (snRNP) code sequence and is essential, together with other components of the survival of motor neurons (SMN) complex, for the biogenesis of snRNPs, the major constituents of spliceosomes. We show that this binding is mediated by Gemin5's WD repeat domain, a common domain not previously known to bind RNA independently. The entire WD repeat domain, comprising 13 WD motifs, is both necessary and sufficient for sequence-specific, high-affinity binding of Gemin5 to its RNA targets. Using an RNA-mediated hydroxyl radical probing method and mass spectrometry, we mapped a discrete region of the WD repeat domain that contacts snRNAs and demonstrated by mutagenesis that specific amino acids in this region are crucial for Gemin5-snRNA binding. The WD repeat domain is thus a previously undescribed RNA binding domain, and we suggest that the presence of WD repeats should be considered as predictive of potential function in RNA binding.

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Figure 1: The WD repeat domain of Gemin5 (G5) specifically binds to RNAs containing a snRNP code sequence.
Figure 2: Mapping of the RNA binding site in the WD repeat domain of Gemin5 (G5).
Figure 3: Mapping of the RNA binding site in the WD repeat domain.
Figure 4: A model of the predicted structure of Gemin5's WD repeat domain.
Figure 5: Site-directed mutagenesis of Gemin5 (G5) and its effect on RNA binding.

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Acknowledgements

We thank the members of our laboratory, especially D. Battle, L. Wan, J. Yong, M. Kasim and I. Younis, for stimulating discussions and helpful comments on this manuscript. We thank S. Pesiridis and K. Lorenz for help with mass spectrometry. This work was supported by the Association Française Contre les Myopathies (AFM). G.D. is funded by the Howard Hughes Medical Institute.

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Authors and Affiliations

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

    Chi-kong Lau, Jennifer L Bachorik & Gideon Dreyfuss

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  1. Chi-kong Lau
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  2. Jennifer L Bachorik
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  3. Gideon Dreyfuss
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Contributions

C-k.L. designed and performed the experiments and wrote the manuscript; J.L.B. performed the experiments; G.D. directed the project, designed experiments and wrote the manuscript.

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Correspondence to Gideon Dreyfuss.

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Lau, Ck., Bachorik, J. & Dreyfuss, G. Gemin5-snRNA interaction reveals an RNA binding function for WD repeat domains. Nat Struct Mol Biol 16, 486–491 (2009). https://doi.org/10.1038/nsmb.1584

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