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The Role of L1 Stalk-tRNA Interaction in the Ribosome Elongation Cycle
Authors:Leonardo G Trabuco  Eduard Schreiner  Peter Cornish  Taekjip Ha  Klaus Schulten
Institution:
  • 1 Beckman Institute for Advanced Science and Technology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • 2 Center for Biophysics and Computational Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • 3 Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • 4 Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA
  • Abstract:The ribosomal L1 stalk is a mobile structure implicated in directing tRNA movement during translocation through the ribosome. This article investigates three aspects of L1 stalk-tRNA interaction. First, by combining data from cryo electron microscopy, X-ray crystallography, and molecular dynamics simulations through the molecular dynamics flexible fitting method, we obtained atomic models of different tRNAs occupying the hybrid P/E state interacting with the L1 stalk. These models confirm the assignment of fluorescence resonance energy transfer states from previous single-molecule investigations of L1 stalk dynamics. Second, the models reconcile how initiator tRNAfMet interacts less strongly with the L1 stalk compared to elongator tRNAPhe, as seen in previous single-molecule experiments. Third, results from a simulation of the entire ribosome in which the L1 stalk is moved from a half-closed conformation to its open conformation are found to support the hypothesis that L1 stalk opening is involved in tRNA release from the ribosome.
    Keywords:rRNA  ribosomal RNA  ASL  anticodon-stem loop  smFRET  single-molecule fluorescence resonance energy transfer  EF-G  elongation factor G  cryo-EM  cryo electron microscopy  MDFF  molecular dynamics flexible fitting  MD  molecular dynamics  RMSF  root-mean-square fluctuation  EF-Tu  elongation factor Tu  FRET  fluorescence resonance energy transfer  PDB  Protein Data Bank  PME  particle mesh Ewald
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