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Related Experiment Videos

Tertiary structure stabilization promotes hairpin ribozyme ligation.

M J Fedor1

  • 1Department of Molecular Biology, The Skaggs Institute for Chemical Biology, The Scripps Research Institute, La Jolla, California 92037, USA. mfedor@scripps.edu

Biochemistry
|August 25, 1999
PubMed
Summary
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The catalytic mechanism of the hairpin ribozyme.

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Hairpin ribozymes with four-way helical junctions mediate intracellular RNA ligation.

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Structure and function of the hairpin ribozyme.

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The kinetic mechanism of the hairpin ribozyme in vivo: influence of RNA helix stability on intracellular cleavage kinetics.

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Kinetics of hairpin ribozyme cleavage in yeast.

RNA (New York, N.Y.)·1997

The hairpin ribozyme

Area of Science:

  • Molecular Biology
  • Biochemistry
  • RNA Catalysis

Background:

  • The hairpin ribozyme is crucial for processing viral satellite RNA replication intermediates in plants.
  • It features two helix-loop-helix segments that associate noncoaxially in its active structure.
  • The hairpin ribozyme in Tobacco Ringspot Virus satellite RNA assembles within a four-way helical junction.

Purpose of the Study:

  • To investigate the impact of a four-way helical junction on hairpin ribozyme structure and function.
  • To analyze the kinetic mechanism changes associated with this structural modification.
  • To explore the relationship between RNA tertiary structure stability and ribozyme activity.

Main Methods:

  • Physical characterization of hairpin ribozyme structures using fluorescence resonance energy transfer (FRET).

Related Experiment Videos

  • Analysis of kinetic mechanisms, including cleavage and ligation reactions.
  • Comparison of hairpin ribozymes with and without a four-way helical junction.

    • Main Results:

    • The four-way helical junction enhances the stability of the folded hairpin ribozyme structure.
    • Ribozymes with the four-way junction exhibit significantly higher affinity for 3' cleavage products.
    • The balance of the reaction shifts towards ligation, indicating enhanced ligase activity.

    Conclusions:

    • Tertiary structure stability, influenced by the four-way helical junction, is a key determinant of hairpin ribozyme proficiency.
    • Enhanced stability of the folded structure contributes to increased product binding affinity.
    • The findings illustrate a direct link between RNA structure and biological function in ribozyme catalysis.