Domain movements of elongation factor eEF2 and the eukaryotic 80S ribosome facilitate tRNA translocation

EMBO J. 2004 Mar 10;23(5):1008-19. doi: 10.1038/sj.emboj.7600102. Epub 2004 Feb 19.

Abstract

An 11.7-A-resolution cryo-EM map of the yeast 80S.eEF2 complex in the presence of the antibiotic sordarin was interpreted in molecular terms, revealing large conformational changes within eEF2 and the 80S ribosome, including a rearrangement of the functionally important ribosomal intersubunit bridges. Sordarin positions domain III of eEF2 so that it can interact with the sarcin-ricin loop of 25S rRNA and protein rpS23 (S12p). This particular conformation explains the inhibitory action of sordarin and suggests that eEF2 is stalled on the 80S ribosome in a conformation that has similarities with the GTPase activation state. A ratchet-like subunit rearrangement (RSR) occurs in the 80S.eEF2.sordarin complex that, in contrast to Escherichia coli 70S ribosomes, is also present in vacant 80S ribosomes. A model is suggested, according to which the RSR is part of a mechanism for moving the tRNAs during the translocation reaction.

Publication types

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

MeSH terms

  • Antifungal Agents / pharmacology
  • Cryoelectron Microscopy
  • Crystallography, X-Ray
  • Indenes
  • Models, Molecular
  • Movement
  • Nucleic Acid Conformation
  • Peptide Elongation Factor 2 / chemistry*
  • Peptide Elongation Factor 2 / metabolism*
  • Peptide Elongation Factor 2 / ultrastructure
  • Protein Binding
  • Protein Structure, Quaternary
  • Protein Structure, Tertiary
  • Protein Subunits / genetics
  • Protein Subunits / metabolism
  • RNA Transport
  • RNA, Transfer / chemistry
  • RNA, Transfer / genetics
  • RNA, Transfer / metabolism*
  • Ribosomes / chemistry*
  • Ribosomes / metabolism*
  • Ribosomes / ultrastructure
  • Saccharomyces cerevisiae / chemistry
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / ultrastructure

Substances

  • Antifungal Agents
  • Indenes
  • Peptide Elongation Factor 2
  • Protein Subunits
  • sordarin
  • RNA, Transfer

Associated data

  • PDB/1S1H
  • PDB/1S1I