Identification of eukaryotic and prokaryotic methylthiotransferase for biosynthesis of 2-methylthio-N6-threonylcarbamoyladenosine in tRNA

Simon Arragain, Samuel K. Handelman, Farhad Forouhar, Fan Yan Wei, Kazuhito Tomizawa, John F. Hunt, Thierry Douki, Marc Fontecave, Etienne Mulliez, Mohamed Atta

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Abstract

Bacterial and eukaryotic transfer RNAs have been shown to contain hypermodified adenosine, 2-methylthio-N6-threonylcarbamoyladenosine, at position 37 (A37) adjacent to the 3′-end of the anticodon, which is essential for efficient and highly accurate protein translation by the ribosome. Using a combination of bioinformatic sequence analysis and in vivo assay coupled to HPLC/MS technique, we have identified, from distinct sequence signatures, two methylthiotransferase (MTTase) subfamilies, designated as MtaB in bacterial cells and e-MtaB in eukaryotic and archaeal cells. Both subfamilies are responsible for the transformation of N6- threonylcarbamoyladenosine into 2-methylthio-N6- threonylcarbamoyladenosine. Recently, a variant within the human CDKAL1 gene belonging to the e-MtaB subfamily was shown to predispose for type 2 diabetes. CDKAL1 is thus the first eukaryotic MTTase identified so far. Using purified preparations of Bacillus subtilis MtaB (YqeV), a CDKAL1 bacterial homolog, we demonstrate that YqeV/CDKAL1 enzymes, as the previously studied MTTases MiaB and RimO, contain two [4Fe-4S] clusters. This work lays the foundation for elucidating the function of CDKAL1.

Original languageEnglish
Pages (from-to)28425-28433
Number of pages9
JournalJournal of Biological Chemistry
Volume285
Issue number37
DOIs
Publication statusPublished - 2010 Sep 10

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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    Arragain, S., Handelman, S. K., Forouhar, F., Wei, F. Y., Tomizawa, K., Hunt, J. F., Douki, T., Fontecave, M., Mulliez, E., & Atta, M. (2010). Identification of eukaryotic and prokaryotic methylthiotransferase for biosynthesis of 2-methylthio-N6-threonylcarbamoyladenosine in tRNA. Journal of Biological Chemistry, 285(37), 28425-28433. https://doi.org/10.1074/jbc.M110.106831