Interaction of the G' domain of elongation factor G and the C-terminal domain of ribosomal protein L7/L12 during translocation as revealed by cryo-EM.

Abstract

During tRNA translocation on the ribosome, an arc-like connection (ALC) is formed between the G' domain of elongation factor G (EF-G) and the L7/L12-stalk base of the large ribosomal subunit in the GDP state. To delineate the boundary of EF-G within the ALC, we tagged an amino acid residue near the tip of the G' domain of EF-G with undecagold, which was then visualized with three-dimensional cryo-electron microscopy (cryo-EM). Two distinct positions for the undecagold, observed in the GTP-state and GDP-state cryo-EM maps of the ribosome bound EF-G, allowed us to determine the movement of the labeled amino acid. Molecular analyses of the cryo-EM maps show: (1) that three structural components, the N-terminal domain of ribosomal protein L11, the C-terminal domain of ribosomal protein L7/L12, and the G' domain of EF-G, participate in formation of the ALC; and (2) that both EF-G and the ribosomal protein L7/L12 undergo large conformational changes to form the ALC.

• Authors: Partha P Datta, Manjuli R Sharma, Li Qi, Joachim Frank, Rajendra K Agrawal
• Journal: Molecular cell (Mol Cell) Vol. 20 Issue 5 Pg. 723-31 (Dec 09 2005) ISSN: 1097-2765 [Print] United States
• PMID: 16337596 (Publication Type: Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, Non-P.H.S.)

Chemical References

• Peptide Elongation Factor G
• Ribosomal Proteins
• ribosomal protein L7-L12
• Gold
• RNA, Transfer

Topics

• Cryoelectron Microscopy (methods)
• Crystallography, X-Ray
• Escherichia coli (chemistry, metabolism)
• Gold (chemistry)
• Hydrolysis
• Models, Molecular
• Peptide Elongation Factor G (chemistry, metabolism)
• Protein Conformation
• Protein Structure, Tertiary
• RNA, Transfer (chemistry, metabolism)
• Ribosomal Proteins (chemistry, metabolism)
• Ribosomes (chemistry, metabolism, ultrastructure)