The ATP-mediated formation of the YgjD–YeaZ–YjeE complex is required for the biosynthesis of tRNA t6A in Escherichia coli

Wenhua Zhang (1) , Bruno Collinet (1) , Ludovic Perrochia (2) , D. Durand (1) , Herman van Tilbeurgh (1)

1 IBBMC - Institut de biochimie et biophysique moléculaire et cellulaire
2 IGM - Institut de génétique et microbiologie [Orsay]

The essential and universal N 6-threonylcarbamoyladenosine (t 6 A) modification at position 37 of ANN-decoding tRNAs plays a pivotal role in translational fidelity through enhancement of the cognate codon recognition and stabilization of the codon–anticodon interaction. In Escherichia coli, the YgjD (TsaD), YeaZ (TsaB), YjeE (TsaE) and YrdC (TsaC) proteins are necessary and sufficient for the in vitro biosyn-thesis of t 6 A, using tRNA, ATP, L-threonine and bicarbonate as substrates. YrdC synthesizes the short-lived L-threonylcarbamoyladenylate (TCA), and YgjD, YeaZ and YjeE cooperate to transfer the L-threonylcarbamoyl-moiety from TCA onto adenosine at position 37 of substrate tRNA. We determined the crystal structure of the heterodimer YgjD–YeaZ at 2.3 ˚ A, revealing the presence of an unexpected molecule of ADP bound at an atypical site situated at the YgjD–YeaZ interface. We further showed that the ATPase activity of YjeE is strongly activated by the YgjD–YeaZ heterodimer. We established by binding experiments and SAXS data analysis that YgjD–YeaZ and YjeE form a compact ternary complex only in presence of ATP. The formation of the ternary YgjD–YeaZ–YjeE complex is required for the in vitro biosynthesis of t 6 A but not its ATPase activity.

Domaines: Biochimie, Biologie Moléculaire

Wenhua Zhang, Bruno Collinet, Ludovic Perrochia, D. Durand, Herman van Tilbeurgh. The ATP-mediated formation of the YgjD–YeaZ–YjeE complex is required for the biosynthesis of tRNA t6A in Escherichia coli. Nucleic Acids Research, 2015, 43 (3), pp.1804-1817. ⟨10.1093/nar/gku1397⟩. ⟨hal-01277988⟩