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PMID: 19229291 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

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

The EMBO journal ·Vol. 28 ·No. 6 ·2009-03-18 ·Pages 755-65

Schuette JC, Murphy FV, Kelley AC, Weir JR, Giesebrecht J, Connell SR, Loerke J, Mielke T, Zhang W, Penczek PA, Ramakrishnan V, Spahn CM

Abstract

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 A. 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.

MeSH Terms
Cryoelectron Microscopy Enzyme Activation Guanosine Diphosphate/chemistry Models, Molecular Peptide Elongation Factor Tu/chemistry,metabolism,ultrastructure Protein Structure, Secondary Pyridones/chemistry RNA, Transfer/chemistry,ultrastructure Ribosomes/chemistry,enzymology,ultrastructure Static Electricity Thermus thermophilus/enzymology
Chemicals
Pyridones Guanosine Diphosphate RNA, Transfer Peptide Elongation Factor Tu mocimycin
Authors & Affiliations
12 authors, click to expand affiliations / ORCID
Schuette Jan-Christian
Institut für Medizinische Physik und Biophysik, Charite-Universitätsmedizin Berlin, Berlin, Germany.
Murphy 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
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
1460-2075
DOI
10.1038/emboj.2009.26
Published
2009-03-18
Epub
2009-00-19
Pages
755-65
Language
English
Region
England
NLM ID
8208664
PMCID
PMC2666022
Subset
IM
Grants
Medical Research Council · MC_U105184332 · United Kingdom
NIGMS NIH HHS · GM 60635 · United States
Wellcome Trust · United Kingdom
NIGMS NIH HHS · R01 GM060635 · United States
NIGMS NIH HHS · GM 67624 · United States
NIGMS NIH HHS · R01 GM067624 · United States
Databases
PDB
3FIC, 3FIN
Corrections
CommentIn
PMID: 19295500
External Links
PubMed source DOI: 10.1038/emboj.2009.26 PMC Full Text View journal details
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