Fluctuations between multiple EF-G-induced chimeric tRNA states during translocation on the ribosome.

Abstract:

:The coupled translocation of transfer RNA and messenger RNA through the ribosome entails large-scale structural rearrangements, including step-wise movements of the tRNAs. Recent structural work has visualized intermediates of translocation induced by elongation factor G (EF-G) with tRNAs trapped in chimeric states with respect to 30S and 50S ribosomal subunits. The functional role of the chimeric states is not known. Here we follow the formation of translocation intermediates by single-molecule fluorescence resonance energy transfer. Using EF-G mutants, a non-hydrolysable GTP analogue, and fusidic acid, we interfere with either translocation or EF-G release from the ribosome and identify several rapidly interconverting chimeric tRNA states on the reaction pathway. EF-G engagement prevents backward transitions early in translocation and increases the fraction of ribosomes that rapidly fluctuate between hybrid, chimeric and posttranslocation states. Thus, the engagement of EF-G alters the energetics of translocation towards a flat energy landscape, thereby promoting forward tRNA movement.

journal_name

Nat Commun

journal_title

Nature communications

authors

Adio S,Senyushkina T,Peske F,Fischer N,Wintermeyer W,Rodnina MV

doi

10.1038/ncomms8442

subject

Has Abstract

pub_date

2015-06-15 00:00:00

pages

7442

issn

2041-1723

pii

ncomms8442

journal_volume

6

pub_type

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