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Related Experiment Videos

Intermolecular cleavage by the newt ribozyme

L Marusic1, E Luzi, G Barsacchi

  • 1Laboratorio di Biologia Cellulare e dello Sviluppo, Università di Pisa, Italy.

European Journal of Biochemistry
|July 1, 1997
PubMed
Summary
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The newt ribozyme

Area of Science:

  • Molecular Biology
  • RNA catalysis
  • Biochemistry

Background:

  • Hammerhead ribozymes are catalytic RNA molecules.
  • The newt ribozyme is found in newt ovaries.
  • Its trans-cleavage activity requires detailed analysis.

Purpose of the Study:

  • To analyze the trans-cleavage activity of the newt hammerhead ribozyme.
  • To investigate the role of sequence modifications in ribozyme function.
  • To determine the influence of ovary cell extract on ribozyme activity.

Main Methods:

  • Synthesized six ribozymes with natural and modified hammerhead sequences.
  • Tested ribozymes with short oligoribonucleotides and long transcripts.
  • Utilized an RNA secondary-structure folding program.

Related Experiment Videos

Main Results:

  • Natural newt ribozymes showed no activity under multiple turnover conditions.
  • Modified ribozymes with altered stems I and III exhibited improved cleavage.
  • RNA folding analysis revealed proper hammerhead structure formation in modified ribozymes.

Conclusions:

  • Natural newt ribozyme stem sequences hinder hammerhead complex assembly in vitro.
  • Sequence modifications in stems I and III are crucial for catalytic activity.
  • Ovary proteins do not enhance the ribozyme's cleavage function.