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-Structure paper
タイトル | 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. |
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ジャーナル・号・ページ | Nucleic Acids Res, Vol. 46, Issue 11, Page 5861-5874, Year 2018 |
掲載日 | 2018年6月20日 |
著者 | Marcus Fislage / Jingji Zhang / Zuben Patrick Brown / Chandra Sekhar Mandava / Suparna Sanyal / Måns Ehrenberg / Joachim Frank / |
PubMed 要旨 | 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 ...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. |
リンク | Nucleic Acids Res / PubMed:29733411 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 3.0 - 3.6 Å |
構造データ | EMDB-8814: 70S ribosome-EF-Tu H84A complex with GppNHp EMDB-8815, PDB-5we6: EMDB-8826: 70S ribosome-EF-Tu H84A complex with GTP and near-cognate tRNA (Complex I) |
化合物 | ChemComp-FME: ChemComp-MG: ChemComp-K: ChemComp-PHE: ChemComp-GNP: ChemComp-HOH: ChemComp-GTP: |
由来 |
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キーワード | RIBOSOME / translation / GTPase |