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Archaeal Elp3 catalyzes tRNA wobble uridine modification at C5 via a radical mechanism

Nature Chemical Biology volume 10, pages 810–812 (2014)Cite this article

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Abstract

Approximately 25% of cytoplasmic tRNAs in eukaryotic organisms have the wobble uridine (U34) modified at C5 through a process that, according to genetic studies, is carried out by the eukaryotic Elongator complex. Here we show that a single archaeal protein, the homolog of the third subunit of the eukaryotic Elongator complex (Elp3), is able to catalyze the same reaction. The mechanism of action by Elp3 described here represents unprecedented chemistry performed on acetyl-CoA.

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Figure 1: In vitro reconstitution of MinElp3 activity.
Figure 2: Chromatographic and spectrometric analyses of the consumption of cofactors and the production of 5′-dA.
Figure 3: Proposed mechanism of the Elp3-catalyzed reaction.

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Acknowledgements

The study was supported by US National Institute of Health (GM107533 to R.H.H.). We thank Y. Fu and C. He (University of Chicago) for providing us with the synthetic cm5U, F. Sun for assistance with ESI-LC/MS, J. Li and E. Oldfield for assistance with EPR and J. Imlay for the initial use of an anaerobic chamber.

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Authors and Affiliations

  1. Department of Biochemistry, University of Illinois at Urbana-Champaign, Urbana, Illinois, USA

    Kiruthika Selvadurai, Pei Wang, Joseph Seimetz & Raven H Huang

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  1. Kiruthika Selvadurai
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  2. Pei Wang
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  3. Joseph Seimetz
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  4. Raven H Huang
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Contributions

R.H.H. conceived the project. K.S. and R.H.H. designed the experiments. K.S. expressed and purified proteins, in vitro transcribed and purified tRNA substrate and performed in vitro reconstitution. K.S. and R.H.H. performed RP-HPLC and ESI-LC/MS analyses. P.W. and J.S. assisted K.S. in the purification of Elp3 and tRNA. R.H.H. wrote the manuscript with input from other authors.

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Correspondence to Raven H Huang.

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The authors declare no competing financial interests.

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Supplementary Results and Supplementary Figures 1–9. (PDF 2528 kb)

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Selvadurai, K., Wang, P., Seimetz, J. et al. Archaeal Elp3 catalyzes tRNA wobble uridine modification at C5 via a radical mechanism. Nat Chem Biol 10, 810–812 (2014). https://doi.org/10.1038/nchembio.1610

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