Cryo-EM shows stages of initial codon selection on the ribosome by aa-tRNA in ternary complex with GTP and the GTPase-deficient EF-TuH84A

Nucleic Acids Res. 2018 Jun 20;46(11):5861-5874. doi: 10.1093/nar/gky346.

Abstract

The GTPase EF-Tu in ternary complex with GTP and aminoacyl-tRNA (aa-tRNA) promotes rapid and accurate delivery of cognate aa-tRNAs to the ribosomal A site. Here we used cryo-EM to study the molecular origins of the accuracy of ribosome-aided recognition of a cognate ternary complex and the accuracy-amplifying role of the monitoring bases A1492, A1493 and G530 of the 16S rRNA. We used the GTPase-deficient EF-Tu variant H84A with native GTP, rather than non-cleavable GTP analogues, to trap a near-cognate ternary complex in high-resolution ribosomal complexes of varying codon-recognition accuracy. We found that ribosome complexes trapped by GTPase-deficicent ternary complex due to the presence of EF-TuH84A or non-cleavable GTP analogues have very similar structures. We further discuss speed and accuracy of initial aa-tRNA selection in terms of conformational changes of aa-tRNA and stepwise activation of the monitoring bases at the decoding center of the ribosome.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Codon*
  • Cryoelectron Microscopy
  • Guanosine Triphosphate / chemistry*
  • Guanosine Triphosphate / metabolism
  • Models, Molecular
  • Mutation
  • Peptide Elongation Factor Tu / chemistry*
  • Peptide Elongation Factor Tu / genetics
  • Peptide Elongation Factor Tu / metabolism
  • RNA, Messenger / chemistry
  • RNA, Ribosomal, 16S / chemistry
  • RNA, Transfer, Amino Acyl / chemistry*
  • Ribosomes / chemistry*

Substances

  • Codon
  • RNA, Messenger
  • RNA, Ribosomal, 16S
  • RNA, Transfer, Amino Acyl
  • Guanosine Triphosphate
  • Peptide Elongation Factor Tu