Multiperspective smFRET reveals rate-determining late intermediates of ribosomal translocation

Michael R. Wasserman, Jose L. Alejo, Roger B. Altman, Scott C. Blanchard

Research output: Contribution to journalArticlepeer-review

90 Scopus citations


Directional translocation of the ribosome through the mRNA open reading frame is a critical determinant of translational fidelity. This process entails a complex interplay of large-scale conformational changes within the actively translating particle, which together coordinate the movement of tRNA and mRNA substrates with respect to the large and small ribosomal subunits. Using pre-steady state, single-molecule fluorescence resonance energy transfer imaging, we tracked the nature and timing of these conformational events within the Escherichia coli ribosome from five structural perspectives. Our investigations revealed direct evidence of structurally and kinetically distinct late intermediates during substrate movement, whose resolution determines the rate of translocation. These steps involve intramolecular events within the EF-G-GDP-bound ribosome, including exaggerated, reversible fluctuations of the small-subunit head domain, which ultimately facilitate peptidyl-tRNA's movement into its final post-translocation position.

Original languageEnglish (US)
Pages (from-to)333-341
Number of pages9
JournalNature Structural and Molecular Biology
Issue number4
StatePublished - Apr 5 2016

Bibliographical note

Funding Information:
This work was supported by the US National Institutes of Health (2R01GM079238 and 5R01GM098859 to S.C.B.).

Publisher Copyright:
© 2016 Nature America, Inc. All rights reserved.


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