title: Biosynthesis of threonylcarbamoyl adenosine (t6A), a universal tRNA nucleoside.
abstract: The anticodon stem-loop (ASL) of transfer RNAs (tRNAs) drives decoding by interacting directly with the mRNA through codon/anticodon pairing. Chemically complex nucleoside modifications found in the ASL at positions 34 or 37 are known to be required for accurate decoding. Although over 100 distinct modifications have been structurally characterized in tRNAs, only a few are universally conserved, among them threonylcarbamoyl adenosine (t(6)A), found at position 37 in the anticodon loop of a subset of tRNA. Structural studies predict an important role for t(6)A in translational fidelity, and in vivo work supports this prediction. Although pioneering work in the 1970s identified the fundamental substrates for t(6)A biosynthesis, the enzymes responsible for its biosynthesis have remained an enigma. We report here the discovery that in bacteria four proteins (YgjD, YrdC, YjeE, and YeaZ) are both necessary and sufficient for t(6)A biosynthesis in vitro. Notably, YrdC and YgjD are members of universally conserved families that were ranked among the top 10 proteins of unknown function in need of functional characterization, while YeaZ and YjeE are specific to bacteria. This latter observation, coupled with the essentiality of all four proteins in bacteria, establishes this pathway as a compelling new target for antimicrobial development.
authors.affiliation: Department of Chemistry, Portland State University, Portland, Oregon 97207, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Deutsch
authors.forename: Basma
authors.initials: B
authors.lastname: El Yacoubi
authors.forename: Valérie
authors.initials: V
authors.lastname: de Crécy-Lagard
authors.forename: Dirk
authors.initials: D
authors.lastname: Iwata-Reuyl
mesh_terms: D000241:Adenosine / Q000737:chemistry*; D000926:Anticodon; D001483:Base Sequence; D002384:Catalysis; D003062:Codon; D019295:Computational Biology / Q000379:methods; D004926:Escherichia coli / Q000378:metabolism / Q000502:physiology*; D029968:Escherichia coli Proteins / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D013058:Mass Spectrometry / Q000379:methods; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D012343:RNA, Transfer / Q000737:chemistry* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry
chemical_list: D000926:Anticodon; D003062:Codon; D029968:Escherichia coli Proteins; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; C512523:YeaZ protein, E coli; C560833:YgjD protein, E coli; C500230:YjeE protein, E coli; C426646:YrdC protein, E coli; D012343:RNA, Transfer; D000241:Adenosine