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A mouse tRNA pseudogene derived from a tRNA(Trp) coding sequence.

R Ammendola1, T Russo, F Cimino

  • 1Istituto di Scienze Biochimiche, II Facoltà di Medicina e Chirurgia, Napoli.

The Italian Journal of Biochemistry
|March 1, 1988
PubMed
Summary
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Researchers identified a mouse transfer RNA (tRNA) pseudogene. Despite a functional-like promoter sequence, mutations prevent its transcription, indicating transcription does not always occur even with a suitable promoter.

Area of Science:

  • Molecular Biology
  • Genetics
  • Gene Regulation

Background:

  • Transfer RNA (tRNA) genes are essential for protein synthesis.
  • Pseudogenes are non-functional copies of genes that can provide insights into evolutionary processes.
  • Mouse tRNA gene clusters are complex and contain both functional and non-functional elements.

Purpose of the Study:

  • To identify and characterize a novel mouse tRNA(Trp) pseudogene.
  • To investigate the potential for transcription of this pseudogene.
  • To understand the molecular mechanisms that lead to pseudogene formation and inactivation.

Main Methods:

  • Oligonucleotide probing using a synthetic 31-mer complementary to the 3' half of tRNA(Trp).
  • Screening of mouse lambda clones containing tRNA gene clusters.

Related Experiment Videos

  • DNA sequencing to analyze the identified gene's homology and identify mutations.
  • In vitro transcription assays using RNA polymerase III.

    • Main Results:

    • A mouse tRNA(Trp) pseudogene was identified through hybridization with a synthetic oligonucleotide.
    • The pseudogene shows significant homology to a chick tRNA(Trp) gene.
    • Two point mutations and a three-base deletion were found, preventing proper transcript folding.
    • A conserved RNA polymerase III promoter sequence was present, but in vitro transcription was not observed.

    Conclusions:

    • The identified mouse tRNA(Trp) sequence is a pseudogene due to inactivating mutations.
    • The presence of a conserved promoter does not guarantee gene transcription.
    • This finding highlights the complex regulation of gene expression and pseudogene evolution.