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AtaT blocks translation initiation by N-acetylation of the initiator tRNAfMet

Nature Chemical Biology volume 13, pages 640–646 (2017)Cite this article

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Abstract

Toxin–antitoxin (TA) loci are prevalent in bacterial genomes. They are suggested to play a central role in dormancy and persister states. Under normal growth conditions, TA toxins are neutralized by their cognate antitoxins, and under stress conditions, toxins are freed and inhibit essential cellular processes using a variety of mechanisms. Here we characterize ataR–ataT, a novel TA system, from enterohemorrhagic Escherichia coli. We show that the toxin AtaT is a GNAT family enzyme that transfers an acetyl group from acetyl coenzyme A to the amine group of the methionyl aminoacyl moiety of initiator tRNA. AtaT specifically modifies Met-tRNAfMet, but no other aminoacyl-tRNAs, including the elongator Met-tRNAMet. We demonstrate that once acetylated, AcMet-tRNAfMet fails to interact with initiation factor-2 (IF2), resulting in disruption of the translation initiation complex. This work reveals a new mechanism of translation inhibition and confirms Met-tRNAfMet as a prime target to efficiently block cell growth.

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Figure 1: The ataT–ataR gene pair constitutes a type II TA system.
Figure 2: AtaT inhibits translation in an AcCoA-dependent manner.
Figure 3: AtaT acetylates tRNAs.
Figure 4: AtaT acetylates the amine group of the methionine charged on the initiator tRNA.
Figure 5: Translation initiation inhibition in vitro and in vivo by AtaT.
Figure 6: Proposed mode of action of AtaT.

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Acknowledgements

D.J. is a PhD fellow at the Fonds National de la Recherche Scientifique (aspirant FNRS). This work was supported by grants from the Fonds Jean Brachet and the Fondation Van Buuren to L.D.; from the FNRS (grant number F.4505.16 MIS), the Fonds d'Encouragement à la Recherche ULB (FER-ULB), the Fonds Jean Brachet and the Fondation Van Buuren to A.G.P.; and from the FNRS (grant number 3.4621.12 FRSM, T.0147.15F PDR and J.0061.16F CDR), the Interuniversity Attraction Poles Program initiated by the Belgian Science Policy Office (MICRODEV), the Fonds Jean Brachet and the Fondation Van Buuren to L.V.M. The authors would like to thank J.M. Sanz for initial work in characterizing the ataR–ataT system.

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Authors and Affiliations

  1. Laboratoire de Génétique et Physiologie Bactérienne, Université Libre de Bruxelles, Gosselies, Belgium

    Dukas Jurėnas & Laurence Van Melderen

  2. Department of Biochemistry and Molecular Biology, Vilnius University Joint Life Sciences Center, Vilnius, Lithuania

    Dukas Jurėnas

  3. Department of Chemistry, Biomolecular and Analytical Mass Spectrometry Group, University of Antwerp, Antwerp, Belgium

    Sneha Chatterjee, Albert Konijnenberg & Frank Sobott

  4. Astbury Centre for Structural Molecular Biology, University of Leeds, Leeds, UK

    Frank Sobott

  5. School of Molecular and Cellular Biology, University of Leeds, Leeds, UK

    Frank Sobott

  6. Laboratoire de Microbiologie, Université Libre de Bruxelles, Gosselies, Belgium

    Louis Droogmans

  7. Laboratoire de Biologie Structurale et Biophysique, Université Libre de Bruxelles, Gosselies, Belgium

    Abel Garcia-Pino

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Contributions

D.J., L.D., A.G.P. and L.V.M. designed research; D.J., A.G.P., A.K. and S.C. performed research; D.J., A.K., S.C., F.S., L.D., A.G.-P. and L.V.M. analyzed data; and D.J., A.G.-P. and L.V.M. wrote the paper.

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Correspondence to Abel Garcia-Pino or Laurence Van Melderen.

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Jurėnas, D., Chatterjee, S., Konijnenberg, A. et al. AtaT blocks translation initiation by N-acetylation of the initiator tRNAfMet. Nat Chem Biol 13, 640–646 (2017). https://doi.org/10.1038/nchembio.2346

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