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Ubiquitin-related modifier Urm1 acts as a sulphur carrier in thiolation of eukaryotic transfer RNA

Nature volume 458, pages 228–232 (2009)Cite this article

Abstract

Ubiquitin-like proteins (UBLs) can change protein function, localization or turnover by covalent attachment to lysine residues1. Although UBLs achieve this conjugation through an intricate enzymatic cascade, their bacterial counterparts MoaD and ThiS function as sulphur carrier proteins2,3. Here we show that Urm1p, the most ancient UBL3, acts as a sulphur carrier in the process of eukaryotic transfer RNA (tRNA) modification, providing a possible evolutionary link between UBL and sulphur transfer. Moreover, we identify Uba4p, Ncs2p, Ncs6p and Yor251cp as components of this conserved pathway. Using in vitro assays, we show that Ncs6p binds to tRNA, whereas Uba4p first adenylates and then directly transfers sulphur onto Urm1p. Finally, functional analysis reveals that the thiolation function of Urm1p is critical to regulate cellular responses to nutrient starvation and oxidative stress conditions, most likely by increasing translation fidelity.

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Figure 1: Urm1p and Uba4p are functionally linked to the elongator complex.
Figure 2: Cytoplasmic tRNAs from urm1 Δ and uba4 Δ cells are not thiolated.
Figure 3: Ncs2p, Ncs6p and Yor251cp function in the URM1 pathway.
Figure 4: Urm1p acts as a sulphur carrier.

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Acknowledgements

We thank C. Rupp for technical support, M. Sohrmann for yeast strains, G. Rabut for the gift of the pGR51 plasmid, G. L. Igloi and K.-H. Altmann for providing APM, S. Mitani for tut-1(tm1297), B. Casado-Fernandez and D. Hilvert for discussion and technical advice, and T. Kurz and J. D. Alfonzo for reading the manuscript. This work was supported by grants from EuroDyna (to S.L.), the Roche Research Foundation (to P.G.A.P.), ETHZ (to S.L. and M.P.), the SNF (to M.P.), the Canadian Institutes of Health Research (to C.B.) and Genome Canada through the Ontario Genomics Institute (to C.B.).

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Author notes

  1. Sebastian Leidel and Patrick G. A. Pedrioli: These authors contributed equally to this work.

Authors and Affiliations

  1. Institute of Biochemistry, ETH Zurich, Schafmattstrasse 18, CH-8093 Zurich, Switzerland,

    Sebastian Leidel, Patrick G. A. Pedrioli, Tamara Bucher & Matthias Peter

  2. Banting and Best Department of Medical Research and Department of Molecular Genetics, University of Toronto, Toronto, Ontario M5S 3E1, Canada,

    Renée Brost, Michael Costanzo & Charles Boone

  3. Institute of Molecular Systems Biology, ETH Zurich, Wolfgang-Pauli-Strasse 16, CH-8093 Zurich, Switzerland,

    Alexander Schmidt & Ruedi Aebersold

  4. Faculty of Science, University of Zurich, Winterthurerstrasse 190, CH-8057 Zurich, Switzerland,

    Ruedi Aebersold

  5. Institute of Systems Biology, 1441 North 34th Street, Seattle, Washington 98103, USA,

    Ruedi Aebersold

  6. Competence Center for Systems Physiology and Metabolic Diseases, Schafmattstrasse 18, CH-8093 Zurich, Switzerland,

    Alexander Schmidt, Ruedi Aebersold & Matthias Peter

  7. Bioinformatics Group, Miltenyi Biotec GmbH, Friedrich-Ebert-Strasse 68, 51429 Bergisch-Gladbach, Germany,

    Kay Hofmann

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Leidel, S., Pedrioli, P., Bucher, T. et al. Ubiquitin-related modifier Urm1 acts as a sulphur carrier in thiolation of eukaryotic transfer RNA. Nature 458, 228–232 (2009). https://doi.org/10.1038/nature07643

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