Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

tRNA selection and kinetic proofreading in translation.

Scott C Blanchard1, Ruben L Gonzalez, Harold D Kim

  • 1Department of Physics and Applied Physics, Stanford University, Stanford, California 94305-4060, USA.

Nature Structural & Molecular Biology
|September 28, 2004
PubMed
Summary

Single-molecule fluorescence resonance energy transfer (smFRET) revealed the stepwise movement of aminoacyl-tRNA (aa-tRNA) into the ribosome. This study elucidates key intermediate states in tRNA selection and kinetic proofreading during protein synthesis.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Dynamic redistribution of eIF4F controls cap-dependent translation initiation.

bioRxiv : the preprint server for biology·2026
Same author

Remodeling of mRNA by eIF4F in human translation initiation.

bioRxiv : the preprint server for biology·2026
Same author

A ribosomal kinetic checkpoint governs selective mRNA recruitment.

bioRxiv : the preprint server for biology·2026
Same author

Visualizing poloidal orientation in DNA minicircles.

Biophysical journal·2026
Same author

Dynamics of Radiation Damage Buildup in Ultrathin Hexagonal Boron Nitride Films under Ion Bombardment.

ACS applied materials & interfaces·2026
Same author

Toward a unifying mechanistic model of eukaryotic translation initiation through integrative single-molecule, structural, and computational insights.

Current opinion in structural biology·2026

Area of Science:

  • Molecular Biology
  • Biochemistry
  • Structural Biology

Background:

  • Ribosome function is central to protein synthesis.
  • Accurate selection and proofreading of aminoacyl-tRNA (aa-tRNA) are critical for fidelity.
  • The precise mechanisms of aa-tRNA selection and kinetic proofreading remain incompletely understood.

Purpose of the Study:

  • To visualize and characterize the stepwise movement of aa-tRNA into the ribosome.
  • To identify and define intermediate conformational states during tRNA selection and proofreading.
  • To elucidate the roles of specific factors and interactions in these processes.

Main Methods:

  • Single-molecule fluorescence resonance energy transfer (smFRET) to monitor aa-tRNA dynamics.
  • Use of antibiotics (e.g., tetracycline) and nonhydrolyzable GTP analogs to trap intermediates.

Related Experiment Videos

  • Observation of distinct FRET states corresponding to different stages of aa-tRNA binding.

    • Main Results:

    • Identified three distinct FRET states: initial codon recognition, GTPase-activated, and fully accommodated.
    • Tetracycline inhibits aa-tRNA progression at the initial codon recognition state.
    • Sarcin-ricin loop cleavage blocks aa-tRNA movement from the GTPase-activated state.
    • Data support a model where correct codon-anticodon recognition drives rotational movement.

    Conclusions:

    • Proposed a mechanistic model for initial selection and kinetic proofreading of aa-tRNA.
    • Demonstrated the utility of single-molecule methods in dissecting complex ribosomal processes.
    • Provided insights into the dynamic conformational changes governing translation fidelity.