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The Elongator subcomplex Elp456 is a hexameric RecA-like ATPase

Nature Structural & Molecular Biology volume 19, pages 314–320 (2012)Cite this article

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Abstract

Elongator was initially described as an RNA polymerase II–associated factor but has since been associated with a broad range of cellular activities. It has also attracted clinical attention because of its role in certain neurodegenerative diseases. Here we describe the crystal structure of the Saccharomyces cerevisiae subcomplex of Elongator proteins 4, 5 and 6 (Elp456). The subunits each show almost identical RecA folds that form a heterohexameric ring-like structure resembling hexameric RecA-like ATPases. This structural finding is supported by different complementary in vitro and in vivo approaches, including the specific binding of the hexameric Elp456 subcomplex to tRNAs in a manner regulated by ATP. Our results support a role of Elongator in tRNA modification, explain the importance of each of the Elp4, Elp5 and Elp6 subunits for complex integrity and suggest a model for the overall architecture of the holo-Elongator complex.

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Figure 1: Hexameric structure of the rElp456 subcomplex.
Figure 2: rElp456-tRNA interaction.
Figure 3: Elp456 is a hexameric ATPase.
Figure 4: ATP regulates tRNA binding.
Figure 5: Integration of Elp456 subcomplex into the holo-Elongator complex.

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Acknowledgements

The authors acknowledge support by the EMBL Proteomics Core Facility and the EMBL Heidelberg Crystallization Platform and access and support at the ESRF beamlines by the EMBL-ESRF Joint Structural Biology Group. We also thank F. Baudin, O. Barabas, C. Petosa, W. Hübner, C. Sachse and G. Male for comments on the manuscript; A.S. Byström (Umeå University) for the gift of yeast strains and plasmids; J.M. Berger (University of California, Berkeley) for recombinant E. coli Rho protein; D. Rentz, B. Bonneau and M. Correa for technical support; D. Lebert and V. Henriot for help in the early stages of this work; M. Argentini and D. Cornu (SICaPS, IMAGIF platform, Gif sur Yvette, France) for mass spectrometry analysis; and the Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) for their assistance. This work was supported by the European Union 6th Framework programs '3D-Repertoire' (LSHG-CT-2005-512028) to C.W.M. and B.S.; and the CNRS and Ligue Contre le Cancer (Equipe Labellisée 2011) to B.S.

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Authors and Affiliations

  1. European Molecular Biology Laboratory (EMBL), Structural and Computational Biology Unit, Heidelberg, Germany

    Sebastian Glatt, Nicholas M I Taylor & Christoph W Müller

  2. Equipe Labellisée La Ligue, Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Centre National de la Recherche Scientifique (CNRS) UMR7104, Inserm, U964, Illkirch, France

    Juliette Létoquart, Céline Faux & Bertrand Séraphin

  3. Université de Strasbourg, Strasbourg, France

    Juliette Létoquart, Céline Faux & Bertrand Séraphin

  4. Centre de Génétique Moléculaire, Gif sur Yvette, France

    Juliette Létoquart, Céline Faux & Bertrand Séraphin

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  1. Sebastian Glatt
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Contributions

S.G. did the biochemical, biophysical and crystallographic analyses; J.L. optimized expression constructs and initial purifications; C.F. carried out coprecipitations, yeast assays and TAP purifications; N.M.I.T. did the electron microscopy analysis; and S.G., B.S. and C.W.M. designed experiments, analyzed the data and wrote the manuscript.

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Correspondence to Bertrand Séraphin or Christoph W Müller.

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Supplementary Figures 1–6, Supplementary Table 1 and Supplementary Methods (PDF 655 kb)

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Glatt, S., Létoquart, J., Faux, C. et al. The Elongator subcomplex Elp456 is a hexameric RecA-like ATPase. Nat Struct Mol Biol 19, 314–320 (2012). https://doi.org/10.1038/nsmb.2234

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