GTPase activation of elongation factor EF-Tu by the ribosome during decoding

We have used single‐particle reconstruction in cryo‐electron microscopy to determine a structure of the Thermus thermophilus ribosome in which the ternary complex of elongation factor Tu (EF‐Tu), tRNA and guanine nucleotide has been trapped on the ribosome using the antibiotic kirromycin. This repre...

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Bibliographic Details
Published in:The EMBO journal 2009-03, Vol.28 (6), p.755-765
Main Authors: Schuette, Jan-Christian, Murphy IV, Frank V, Kelley, Ann C, Weir, John R, Giesebrecht, Jan, Connell, Sean R, Loerke, Justus, Mielke, Thorsten, Zhang, Wei, Penczek, Pawel A, Ramakrishnan, V, Spahn, Christian M T
Format: Article
Language:English
Subjects:
Cellular biology
cryo-electron microscopy
Cryoelectron Microscopy
Electron microscopy
elongation factor
EMBO31
EMBO40
Enzyme Activation
GTPase
Guanosine Diphosphate - chemistry
Hydrolysis
Models, Molecular
Molecular biology
Peptide Elongation Factor Tu - chemistry
Peptide Elongation Factor Tu - metabolism
Peptide Elongation Factor Tu - ultrastructure
Protein Structure, Secondary
Proteins
Pyridones - chemistry
Ribonucleic acid
ribosome
Ribosomes - chemistry
Ribosomes - enzymology
Ribosomes - ultrastructure
RNA
RNA, Transfer - chemistry
RNA, Transfer - ultrastructure
Signal transduction
Static Electricity
Thermus thermophilus - enzymology
translation
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Summary:We have used single‐particle reconstruction in cryo‐electron microscopy to determine a structure of the Thermus thermophilus ribosome in which the ternary complex of elongation factor Tu (EF‐Tu), tRNA and guanine nucleotide has been trapped on the ribosome using the antibiotic kirromycin. This represents the state in the decoding process just after codon recognition by tRNA and the resulting GTP hydrolysis by EF‐Tu, but before the release of EF‐Tu from the ribosome. Progress in sample purification and image processing made it possible to reach a resolution of 6.4 Å. Secondary structure elements in tRNA, EF‐Tu and the ribosome, and even GDP and kirromycin, could all be visualized directly. The structure reveals a complex conformational rearrangement of the tRNA in the A/T state and the interactions with the functionally important switch regions of EF‐Tu crucial to GTP hydrolysis. Thus, the structure provides insights into the molecular mechanism of signalling codon recognition from the decoding centre of the 30S subunit to the GTPase centre of EF‐Tu.
ISSN:0261-4189
1460-2075
DOI:10.1038/emboj.2009.26