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    A new tRNA intermediate revealed on the ribosome during EF4-mediated back-translocation

    Connell, S.R. and Topf, Maya and Qin, Y. and Wilson, D.N. and Mielke, T. and Fucini, P. and Nierhaus, K.H. and Spahn, C.M.T. (2008) A new tRNA intermediate revealed on the ribosome during EF4-mediated back-translocation. Nature Structural & Molecular Biology 15 , pp. 910-915. ISSN 1545-9985.

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    Abstract

    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.

    Metadata

    Item Type: Article
    School: Birkbeck Schools and Departments > School of Science > Biological Sciences
    Research Centre: Structural Molecular Biology, Institute of (ISMB)
    Depositing User: Administrator
    Date Deposited: 04 Aug 2010 14:09
    Last Modified: 06 Dec 2016 10:35
    URI: http://eprints.bbk.ac.uk/id/eprint/1140

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