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TitleYeast translation elongation factor eEF3 promotes late stages of tRNA translocation.
Journal, issue, pagesEMBO J, Vol. 40, Issue 6, Page e106449, Year 2021
Publish dateMar 15, 2021
AuthorsNamit Ranjan / Agnieszka A Pochopien / Colin Chih-Chien Wu / Bertrand Beckert / Sandra Blanchet / Rachel Green / Marina V Rodnina / Daniel N Wilson /
PubMed AbstractIn addition to the conserved translation elongation factors eEF1A and eEF2, fungi require a third essential elongation factor, eEF3. While eEF3 has been implicated in tRNA binding and release at the ...In addition to the conserved translation elongation factors eEF1A and eEF2, fungi require a third essential elongation factor, eEF3. While eEF3 has been implicated in tRNA binding and release at the ribosomal A and E sites, its exact mechanism of action is unclear. Here, we show that eEF3 acts at the mRNA-tRNA translocation step by promoting the dissociation of the tRNA from the E site, but independent of aminoacyl-tRNA recruitment to the A site. Depletion of eEF3 in vivo leads to a general slowdown in translation elongation due to accumulation of ribosomes with an occupied A site. Cryo-EM analysis of native eEF3-ribosome complexes shows that eEF3 facilitates late steps of translocation by favoring non-rotated ribosomal states, as well as by opening the L1 stalk to release the E-site tRNA. Additionally, our analysis provides structural insights into novel translation elongation states, enabling presentation of a revised yeast translation elongation cycle.
External linksEMBO J / PubMed:33555093 / PubMed Central
MethodsEM (single particle)
Resolution3.3 - 4.2 Å
Structure data

EMDB-12059:
Yeast 80S ribosome bound to eEF3 and A/A- and P/P-tRNAs (PRE-1)
Method: EM (single particle) / Resolution: 3.5 Å

EMDB-12061:
Yeast 80S ribosome bound to eEF3 and A/A- and P/E-tRNAs (PRE-2).
Method: EM (single particle) / Resolution: 4.0 Å

EMDB-12062:
Yeast 80S ribosome bound to eEF3 and P/P- and E/E-tRNAs (POST-2)
Method: EM (single particle) / Resolution: 3.9 Å

EMDB-12064:
Yeast 80S ribosome bound to eEF3 and P/P-tRNA (POST-3)
Method: EM (single particle) / Resolution: 3.8 Å

EMDB-12065:
Yeast 80S ribosome with bound A/P- and P/E-tRNAs (PRE-4)
Method: EM (single particle) / Resolution: 3.3 Å

EMDB-12074:
Yeast 80S ribosome bound to eEF2 and ap/P- and pe/E-tRNAs (POST-1).
Method: EM (single particle) / Resolution: 4.1 Å

EMDB-12075:
Yeast 80S ribosome with bound A/A- and P/E-tRNAs (PRE-3).
Method: EM (single particle) / Resolution: 4.2 Å

Source
  • Saccharomyces cerevisiae (brewer's yeast)

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