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EF-G Mutations Reveal Correlation between Power Stroke and Translocation Fidelity in Protein Synthesis.

Yanjun Chen1, Jacob H Steele2, Shoujun Xu1

  • 1Department of Chemistry, University of Houston, Houston, TX 77204, USA.

Biorxiv : the Preprint Server for Biology
|July 16, 2025
PubMed
Summary

Quantum sensing reveals how mutations in elongation factor G (EF-G) affect ribosome translocation. Altering EF-G

Keywords:
elongation factor Gforce spectroscopypower strokequantum sensingribosome translocation

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Area of Science:

  • Molecular Biology
  • Biophysics
  • Quantum Sensing

Background:

  • Ribosome translocation is a fundamental process in protein synthesis, moving along mRNA in three-nucleotide steps.
  • Elongation factor G (EF-G) catalyzes ribosome translocation, undergoing large conformational changes that generate mechanical force (power stroke).
  • The precise quantification of EF-G's power stroke and its role in translocation fidelity remain incompletely understood.

Purpose of the Study:

  • To investigate the relationship between EF-G's power stroke and ribosome translocation fidelity using quantum sensing.
  • To analyze the mechanical force generated by mutated EF-G variants and their impact on mRNA translocation accuracy.

Main Methods:

  • Expression and characterization of two EF-G mutants: H584K and Q508K.
  • Application of quantum sensing for combined measurements of EF-G power stroke.
  • Analysis of ribosome translocation steps and frameshifting induced by EF-G mutants.

Main Results:

  • The H584K EF-G mutant, interacting with the codon-anticodon minihelix, showed a reduced power stroke (60 ± 6 pN) and induced -1 frameshifting (2-nt translocation).
  • The Q508K EF-G mutant, interacting outside the minihelix, exhibited a near-wild-type power stroke (89 ± 11 pN) and maintained 3-nt translocation.
  • Direct correlation observed between EF-G power stroke magnitude and translocation fidelity.

Conclusions:

  • EF-G's force projection and pivotal interaction points are critical for maintaining accurate ribosome translocation.
  • Reduced power stroke in EF-G mutants leads to frameshifting errors during protein synthesis.
  • Findings suggest EF-G's mechanical force influences the kinetic energy barrier of translocation, ensuring fidelity.