Abstract
Eukaryotic cells have several quality control pathways that rely on translation to detect and degrade defective RNAs. Dom34 and Hbs1 are two proteins that are related to translation termination factors and are involved in no-go decay (NGD) and nonfunctional 18S ribosomal RNA (rRNA) decay (18S NRD) pathways that eliminate RNAs that cause strong ribosomal stalls. Here we present the structure of Hbs1 with and without GDP and a low-resolution model of the Dom34–Hbs1 complex. This complex mimics complexes of the elongation factor and transfer RNA or of the translation termination factors eRF1 and eRF3, supporting the idea that it binds to the ribosomal A-site. We show that nucleotide binding by Hbs1 is essential for NGD and 18S NRD. Mutations in Hbs1 that disrupted the interaction between Dom34 and Hbs1 strongly impaired NGD but had almost no effect on 18S NRD. Hence, NGD and 18S NRD could be genetically uncoupled, suggesting that mRNA and rRNA in a stalled translation complex may not always be degraded simultaneously.
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Acknowledgements
We thank K. Blondeau, B. Faivre, J. Cicolari, D. Lebert, Y. Billier, B. Bonneau and D. Rentz for technical assistance; M. Moore (U. Mass. Medical School) and R. Parker (University of Arizona) for plasmid gifts; and P. Vachette for discussions. Supported by the Agence Nationale pour la Recherche (ANR-06-BLAN-0075-02 and ANR-07-BLAN-0093), the Association Française contre les Myopathies (AFM), La Ligue contre le Cancer Equipe Labellisée 2008, CNRS, the ESF EUROCORES RNA Quality and the EU '3D-Repertoire' program (LSHG-CT-2005-512028). J.H. and A.M.G.v.d.E. hold predoctoral grants from the Université Paris-Sud 11 and Université de Strasbourg, respectively. M.E.G. is supported by the Spanish Ministry of Science and Innovation. We acknowledge SOLEIL for provision of synchrotron radiation facilities and thank A. Thompson and J. Perez for assistance with beamlines Proxima-1 and SWING, respectively.
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J.H., A.M.G.v.d.E., M.G. and B.S. designed experiments. J.H., A.M.G.v.d.E., N.L., D.D., F.L., M.N. and M.E.G. performed experiments. J.H., A.M.G.v.d.E., D.D., H.v.T., M.G. and B.S. analyzed data and wrote the paper.
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van den Elzen, A., Henri, J., Lazar, N. et al. Dissection of Dom34–Hbs1 reveals independent functions in two RNA quality control pathways. Nat Struct Mol Biol 17, 1446–1452 (2010). https://doi.org/10.1038/nsmb.1963
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DOI: https://doi.org/10.1038/nsmb.1963
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