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Related Experiment Videos

Ligand-induced conformational changes observed in single RNA molecules.

T Ha1, X Zhuang, H D Kim

  • 1Department of Physics, Stanford University, Stanford, CA 94305, USA.

Proceedings of the National Academy of Sciences of the United States of America
|August 4, 1999
PubMed
Summary

We demonstrate using fluorescence resonance energy transfer (FRET) to track single RNA molecule motion during conformational changes. This method also functions as the world's smallest magnesium ion (Mg2+) meter with millisecond resolution.

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Area of Science:

  • Biophysics
  • Molecular Biology
  • Biochemistry

Background:

  • Conformational changes in RNA are crucial for biological function.
  • Understanding protein-RNA interactions requires studying molecular dynamics.
  • Existing bulk assays lack single-molecule resolution.

Purpose of the Study:

  • To demonstrate fluorescence resonance energy transfer (FRET) for tracking single RNA molecule conformational changes.
  • To investigate RNA conformational dynamics induced by protein binding or ion concentration.
  • To develop a novel method for real-time monitoring of molecular interactions.

Main Methods:

  • Utilized single-molecule fluorescence resonance energy transfer (smFRET).
  • Studied three-helix junction RNA molecules with attached dye probes.

Related Experiment Videos

  • Induced conformational changes using ribosomal protein S15 and magnesium ions (Mg2+).
  • Monitored transitions between open and folded RNA states.

    • Main Results:

    • Successfully tracked single RNA molecule conformational changes using smFRET.
    • Observed behavior consistent with bulk assays, validating the smFRET approach.
    • Detected an anomalously broad distribution of RNA conformations at intermediate Mg2+ concentrations.
    • Demonstrated real-time monitoring of RNA response to changing Mg2+ concentrations with 20-ms resolution.

    Conclusions:

    • smFRET is a viable technique for studying single RNA molecule dynamics and protein-nucleic acid interactions.
    • Intermediate Mg2+ concentrations reveal heterogeneity in RNA folding.
    • Single RNA molecules can act as highly sensitive, miniature Mg2+ sensors.