Agirrezabala, Xabier; Schreiner, Eduard; Trabuco, Leonardo G.; Lei, Jianlin; Ortiz-Meoz, Rodrigo F.; Schulten, Klaus; Green, Rachel; Frank, Joachim
Structural insights into cognate versus near-cognate discrimination during decoding
EMBO JOURNAL, 30:1497-1507, APR 20 2011

The structural basis of the tRNA selection process is investigated by cryo-electron microscopy of ribosomes programmed with UGA codons and incubated with ternary complex (TC) containing the near-cognate Trp-tRNA(Trp) in the presence of kirromycin. Going through more than 350 000 images and employing image classification procedures, we find similar to 8% in which the TC is bound to the ribosome. The reconstructed 3D map provides a means to characterize the arrangement of the near-cognate aa-tRNA with respect to elongation factor Tu (EF-Tu) and the ribosome, as well as the domain movements of the ribosome. One of the interesting findings is that near-cognate tRNA's acceptor stem region is flexible and CCA end becomes disordered. The data bring direct structural insights into the induced-fit mechanism of decoding by the ribosome, as the analysis of the interactions between small and large ribosomal subunit, aa-tRNA and EF-Tu and comparison with the cognate case (UGG codon) offers clues on how the conformational signals conveyed to the GTPase differ in the two cases. The EMBO Journal (2011) 30, 1497-1507. doi:10.1038/emboj.2011.58; Published online 4 March 2011 Subject Categories: proteins; structural biology

DOI:10.1038/emboj.2011.58

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