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Document Version Version Comment Date Status
407108.0 [No comment] 04.03.2009 13:33 Released

ID: 407108.0, MPI für molekulare Genetik / Department of Vertebrate Genomics
A new tRNA intermediate revealed on the ribosome during EF4-mediated back-translocation
Authors:Connell, Sean R.; Topf, Maya; Qin, Yan; Wilson, Daniel N.; Mielke, Thorsten; Fucini, Paola; Nierhaus, Knud H.; Spahn, Christian M. T.
Date of Publication (YYYY-MM-DD):2008-09
Title of Journal:Nature Structural & Molecular Biology
Journal Abbrev.:Nat Struct Mol Biol
Issue / Number:9
Start Page:910
End Page:915
Copyright:© 2009 Nature Publishing Group
Review Status:not specified
Audience:Experts Only
Abstract / Description:EF4 (LepA) is an almost universally conserved translational GTPase in eubacteria. It seems to be essential under environmental stress conditions and has previously been shown to back-translocate the tRNAs on the ribosome, thereby reverting the canonical translocation reaction. In the current work, EF4 was directly visualized in the process of back-translocating tRNAs by single-particle cryo-EM. Using flexible fitting methods, we built a model of ribosome-bound EF4 based on the cryo-EM map and a recently published unbound EF4 X-ray structure. The cryo-EM map establishes EF4 as a noncanonical elongation factor that interacts not only with the elongating ribosome, but also with the back-translocated tRNA in the A-site region, which is present in a previously unseen, intermediate state and deviates markedly from the position of a canonical A-tRNA. Our results, therefore, provide insight into the underlying structural principles governing back-translocation.
Comment of the Author/Creator:E-mail:
External Publication Status:published
Document Type:Article
Communicated by:Hans Lehrach
Affiliations:MPI für molekulare Genetik
External Affiliations:Institut für Medizinische Physik und Biophysik, Charite–Universitätsmedizin Berlin, Ziegelstrasse 5-9, 10117-Berlin, Germany
Cluster of Excellence for Macromolecular Complexes, Institut für Organische Chemie und Chemische Biologie, J.W. Goethe Universität, Max-von-Laue Strasse 7, 60438 Frankfurt am Main, Germany
Institute of Structural and Molecular Biology, School of Crystallography, Birkbeck College, Malet Street, London WC1E 7HX, UK
Gene Center and Department for Chemistry and Biochemistry, University of Munich, Feodor-Lynen-Strasse 25, 81377 Munich, Germany
Munich Center for Integrated Protein Science (CiPSM), University of Munich, 81377 Munich, Germany
UltraStrukturNetzwerk, Max Planck Institute für Molekulare Genetik, Ihnestrasse 73, 14195-Berlin, Germany
National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 15 Datun Road, Chaoyang District, Beijing 100101, China