Single-Molecule Structural Dynamics of EF-G−Ribosome Interaction during Translocation
Citations Over TimeTop 10% of 2007 papers
Abstract
EF-G catalyzes translocation of mRNA and tRNAs within the ribosome during protein synthesis. Detection of structural states in the reaction sequence that are not highly populated can be facilitated by studying the process one molecule at a time. Here we present single-molecule studies of translocation showing that, for ribosomes engaged in poly(Phe) synthesis, fluorescence resonance energy transfer (FRET) between the G‘ domain of EF-G and the N-terminal domain of ribosomal protein L11 occurs within two rapidly interconverting states, having FRET efficiencies of 0.3 and 0.6. The antibiotic fusidic acid increases the population of the 0.6 state, indicating that it traps the ribosome·EF-G complex in a preexisting conformation formed during translation. Only the 0.3 state is observed when poly(Phe) synthesis is prevented by omission of EF-Tu, or in studies on vacant ribosomes. These results suggest that the 0.6 state results from the conformational lability of unlocked ribosomes formed during translocation. An idling state, possibly pertinent to regulation of protein synthesis, is detected in some ribosomes in the poly(Phe) system.
Related Papers
- → Blasticidin S inhibits translation by trapping deformed tRNA on the ribosome(2013)112 cited
- → Role of the ribosomal protein L27 revealed by single‐molecule FRET study(2012)10 cited
- → The kinetics of the translation of messenger RNA into protein(1967)11 cited
- → Substructures in Ribosomes of E. coli ? A Hypothesis for the Attachment of Transfer RNAs(1969)11 cited
- Ribosomes and subunits from Escherichia coli studied by asymmetrical flow field-flow fractionation(1998)