TCB Publications - Abstract

Xabier Agirrezabala, Hstau Liao, Eduard Schreiner, Jie Fu, Rodrigo Ortiz-Meoz, Klaus Schulten, Rachel Green, and Joachim Frank. Structural characterization of mRNA-tRNA translocation intermediates. Proceedings of the National Academy of Sciences, USA, 109:6094-6099, 2012. (PMC: 3340995)

AGIR2012 Analysis of a wild-type E. coli pre-translocational sample competent for unimpaired forward translocation by cryo-EM, classification, and single-particle reconstruction has revealed the presence of previously unseen intermediate substates of the bacterial ribosome during the first phase of translocation, characterized by intermediate intersubunit rotations, L1 stalk positions, and tRNA configurations. Furthermore, we have described the domain rearrangements in quantitative terms, which has allowed us to define the processivity and coordination of the conformational reorganization of the ribosome, along with the associated changes in tRNA ribosome-binding configuration. The observed occupancies of substates also allowed a profile of the free-energy landscape to be reconstructed. The results are consistent with the view of the ribosome as a molecular machine employing Brownian motion to reach a functionally productive state (“macrostate II”) via a series of substates with incremental changes in conformation.

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