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Cryo-EM structures of Arx1 and maturation factors Rei1 and Jjj1 bound to the 60S ribosomal subunit

Nature Structural & Molecular Biology volume 19, pages 1228–1233 (2012)Cite this article

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Abstract

Eukaryotic ribosome biogenesis requires many protein factors that facilitate the assembly, nuclear export and final maturation of 40S and 60S particles. We have biochemically characterized ribosomal complexes of the yeast 60S-biogenesis factor Arx1 and late-maturation factors Rei1 and Jjj1 and determined their cryo-EM structures. Arx1 was visualized bound to the 60S subunit together with Rei1, at 8.1-Å resolution, to reveal the molecular details of Arx1 binding whereby Arx1 arrests the eukaryotic-specific rRNA expansion segment 27 near the polypeptide tunnel exit. Rei1 and Jjj1, which have been implicated in Arx1 recycling, bind in the vicinity of Arx1 and form a network of interactions. We suggest that, in addition to the role of Arx1 during pre-60S nuclear export, the binding of Arx1 conformationally locks the pre-60S subunit and inhibits the premature association of nascent chain–processing factors to the polypeptide tunnel exit.

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Figure 1: Biochemical characterization of the complexes of Rei1, Arx1 and Jjj1 with the 60S ribosomal subunit.
Figure 2: Cryo-EM reconstruction of the 60S–Arx1–Rei1 complex.
Figure 3: Detailed interpretation of the structure of 60S-bound Arx1.
Figure 4: Rei1 in the 60S–Arx1–Rei1 complex.
Figure 5: Cryo-EM reconstruction of the 60S–Arx1–Rei1–Jjj1 complex.
Figure 6: Comparison of the binding sites of ribosome-biogenesis factors and nascent chain–processing factors near the polypeptide tunnel exit.
Figure 7: Schematic model of the Arx1 release process.

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Acknowledgements

We thank M. Leibundgut for critically reading this manuscript and E. Deuerling and T. Maier for discussions. We thank Electron Microscopy ETH Zurich (EMEZ) for data collection and P. Tittmann for support. This work was supported by the Swiss National Science Foundation (SNSF), the National Center of Excellence in Research (NCCR) Structural Biology program of the SNSF, and European Research Council (ERC) grant 250071 under the European Community's Seventh Framework Program (to N.B.).

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  1. Christian Montellese

    Present address: Current address: Institute of Biochemistry, ETH Zürich, Zürich, Switzerland.,

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  1. Institute of Molecular Biology and Biophysics, Eidgenössische Technische Hochschule (ETH) Zürich, Zürich, Switzerland

    Basil J Greber, Daniel Boehringer, Christian Montellese & Nenad Ban

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Contributions

N.B., D.B. and B.J.G. conceived of the study. B.J.G. and C.M. performed biochemistry and sample preparation on the 60S-biogenesis factors and their complexes. B.J.G., D.B. and C.M. acquired cryo-EM data. B.J.G. calculated the cryo-EM structures with the help of D.B., and N.B., D.B. and B.J.G. interpreted the cryo-EM structures and wrote the manuscript.

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Correspondence to Nenad Ban.

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Greber, B., Boehringer, D., Montellese, C. et al. Cryo-EM structures of Arx1 and maturation factors Rei1 and Jjj1 bound to the 60S ribosomal subunit. Nat Struct Mol Biol 19, 1228–1233 (2012). https://doi.org/10.1038/nsmb.2425

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