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Crystal structure of the ribosome recycling factor bound to the ribosome

Nature Structural & Molecular Biology volume 14, pages 733–737 (2007)Cite this article

Abstract

In bacteria, disassembly of the ribosome at the end of translation is facilitated by an essential protein factor termed ribosome recycling factor (RRF), which works in concert with elongation factor G. Here we describe the crystal structure of the Thermus thermophilus RRF bound to a 70S ribosomal complex containing a stop codon in the A site, a transfer RNA anticodon stem-loop in the P site and tRNAfMet in the E site. The work demonstrates that structures of translation factors bound to 70S ribosomes can be determined at reasonably high resolution. Contrary to earlier reports, we did not observe any RRF-induced changes in bridges connecting the two subunits. This suggests that such changes are not a direct requirement for or consequence of RRF binding but possibly arise from the subsequent stabilization of a hybrid state of the ribosome.

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Figure 1: Domain organization of T. thermophilus RRF in the ribosome4,5,6,7.
Figure 2: RRF orientation in the ribosome.
Figure 3: RRF contacts the 70S ribosome.

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Acknowledgements

We thank F. Murphy for help with data collection during optimization of crystallization conditions, M. MacDonald for help with screening at beamline 14.1 at the Synchrotron Radiation Source, and R. Ravelli, J. McCarthy and G. Leonard at beamlines ID14-1, ID14-3 and ID14-4 at the ESRF for help with screening and data collection. This work was supported by the Medical Research Council (UK), the US National Institutes of Health and the Agouron Institute, and fellowships from the Austrian Academy of Sciences (A.W.), the Boehringer Ingelheim Fonds (S.P.), the American Cancer Society (C.M.D.) and the Wenner-Gren foundation (M.S.).

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Author notes

  1. Maria Selmer

    Present address: Present address: Department of Cell and Molecular Biology, Uppsala University, Biomedical Center, Box 596, SE-75124 Uppsala, Sweden.,

  2. Sabine Petry, Christine M Dunham and Maria Selmer: These authors contributed equally to this work.

Authors and Affiliations

  1. MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 0QH, UK

    Albert Weixlbaumer, Sabine Petry, Christine M Dunham, Maria Selmer, Ann C Kelley & V Ramakrishnan

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  1. Albert Weixlbaumer
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Contributions

A.W. crystallized the complex with RRF, collected and analyzed the data and wrote the paper. S.P. and C.M.D. helped with optimization of mRNA constructs, crystallization and cryoprotection conditions, helped with data collection, initial processing and model building, and gave feedback on the manuscript. M.S. cloned, expressed and purified T. thermophilus RRF, suggested crystallization of its complex with the ribosome and provided feedback on the manuscript. A.C.K. purified ribosomes and tRNA. V.R. oversaw the work and helped with writing the paper.

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Correspondence to V Ramakrishnan.

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Weixlbaumer, A., Petry, S., Dunham, C. et al. Crystal structure of the ribosome recycling factor bound to the ribosome. Nat Struct Mol Biol 14, 733–737 (2007). https://doi.org/10.1038/nsmb1282

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