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Structure of the Dom34–Hbs1 complex and implications for no-go decay

Nature Structural & Molecular Biology volume 17, pages 1233–1240 (2010)Cite this article

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Abstract

No-go decay (NGD) targets mRNAs with stalls in translation elongation for endonucleolytic cleavage in a process involving the Dom34 and Hbs1 proteins. The crystal structure of a Schizosaccharomyces pombe Dom34–Hbs1 complex reveals an overall shape similar to that of eRF1–eRF3–GTP and EF-Tu–tRNA–GDPNP. Similarly to eRF1 and GTP binding to eRF3, Dom34 and GTP bind to Hbs1 with strong cooperativity, and Dom34 acts as a GTP-dissociation inhibitor (GDI). A marked conformational change in Dom34 occurs upon binding to Hbs1, leading Dom34 to resemble a portion of a tRNA and to position a conserved basic region in a position expected to be near the peptidyl transferase center. These results support the idea that the Dom34–Hbs1 complex functions to terminate translation and thereby commit mRNAs to NGD. Consistent with this role, NGD at runs of arginine codons, which cause a strong block to elongation, is independent of the Dom34–Hbs1 complex.

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Figure 1: The overall structure of the Dom34–Hbs1 complex.
Figure 2: Comparison of Dom34–Hbs1 with EF-Tu–tRNA–GDPNP and eRF1–eRF3–GTP.
Figure 3: Comparison of Hbs1 in complex.
Figure 4: Conformational changes of Dom34.
Figure 5: The Dom34–Hbs1 interface.
Figure 6: GTP and Dom34 bind to Hbs1 cooperatively and Dom34 acts as a GDI.
Figure 7: NGD at CGA codons is independent of the Dom34–Hbs1 complex.

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Acknowledgements

We would like to thank the beamline scientists at ID23-1 (European Synchrotron Radiation Facility (ESRF), France) for assistance and access to synchrotron radiation facilities. This work is financially supported by the Biomedical Research Council of A*STAR (Agency for Science, Technology and Research) (H.S.), European Regional Development Fund through the Center of Excellence in Chemical Biology (V.H.) and by the Howard Hughes Medical Institute (R.P.).

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Authors and Affiliations

  1. Laboratory of Macromolecular Structure, Institute of Molecular and Cell Biology, Proteos, Singapore

    Liming Chen, Zhihong Cheng, Meng Kiat Lim & Haiwei Song

  2. Department of Molecular and Cellular Biology and Howard Hughes Medical Institute, University of Arizona, Tucson, Arizona, USA

    Denise Muhlrad & Roy Parker

  3. Department of Biomedical Technology, Institute of Technology, University of Tartu, Tartu, Estonia

    Vasili Hauryliuk & Viktoriya Shyp

  4. Department of Biological Sciences, National University of Singapore, Singapore

    Haiwei Song

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Contributions

L.C. solved the crystal structure and performed mutagenesis studies; D.M. carried out NGD assays; V.H. and V.S. performed kinetics assays; Z.C. and M.K.L. carried out cloning and initial purification of Hbs1 and Dom34 proteins; R.P. and H.S. designed the studies, interpreted data and wrote the paper with contributions from L.C. and V.H.

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Correspondence to Roy Parker or Haiwei Song.

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Chen, L., Muhlrad, D., Hauryliuk, V. et al. Structure of the Dom34–Hbs1 complex and implications for no-go decay. Nat Struct Mol Biol 17, 1233–1240 (2010). https://doi.org/10.1038/nsmb.1922

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