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Related Experiment Videos

Tetrahymena ribozyme disrupts rRNA processing in yeast

L Good1, S A Elela, R N Nazar

  • 1Department of Molecular Biology and Genetics, University of Guelph, Ontario, Canada.

The Journal of Biological Chemistry
|September 2, 1994
PubMed
Summary

The Tetrahymena thermophila intervening sequence (IVS) can disrupt ribosomal RNA processing when inserted into the Schizosaccharomyces pombe nuclear genome. This suggests evolutionary pressures may have influenced ribozyme accommodation in Tetrahymena.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • The intervening sequence (IVS) of Tetrahymena thermophila is a group I intron found in nucleolar DNA, interrupting a conserved RNA structure in the large ribosomal subunit.
  • Unlike most group I introns located in mitochondrial and chloroplast DNA, this IVS resides in nuclear DNA, an unusual genomic location.

Purpose of the Study:

  • To investigate the functional impact of inserting the Tetrahymena IVS into a homologous position within the nuclear ribosomal DNA (rDNA) of Schizosaccharomyces pombe.
  • To determine if the Tetrahymena IVS can interfere with rRNA processing in a different nuclear genome.

Main Methods:

  • Introduction of the Tetrahymena IVS into a cloned Schizosaccharomyces pombe ribosomal gene.
  • Expression of the mutant rDNA in vivo within Schizosaccharomyces pombe.

Related Experiment Videos

  • RNA analysis techniques to assess rRNA processing and mature rRNA formation.
  • Hybridization analysis to confirm RNA splicing and free ribozyme presence.

    • Main Results:

    • Mature 5.8 S ribosomal RNA (rRNA) was not produced from the mutant gene transcript.
    • A significant reduction in the quantity of 27 S precursor rRNA (nRNA) was observed.
    • Despite processing interference, RNA splicing still occurred, and functional free ribozyme was detected.

    Conclusions:

    • The Tetrahymena IVS sequence has the capacity to interfere with rRNA processing in heterologous nuclear genomes.
    • The unique nuclear location and amplification of the Tetrahymena rDNA repeat may have necessitated the evolution of mechanisms to accommodate its ribozyme.