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Related Experiment Videos

Tissue-specific permutations of self-cleaving newt satellite-2 transcripts.

L M Epstein1, S R Coats

  • 1Department of Biological Science, Florida State University, Tallahassee 32306.

Gene
|November 15, 1991
PubMed
Summary

Newt satellite-2 (sat2) DNA transcripts show self-catalyzed cleavage in vivo, a hammerhead (HH) motif function previously unseen in animal cells. This finding reveals a novel RNA processing mechanism in eukaryotes.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Satellite-2 (sat2) DNA transcripts in newts contain a hammerhead (HH) motif.
  • Hammerhead motifs are known to mediate self-catalyzed RNA cleavage in plant viruses.
  • The in vivo occurrence and function of HH-mediated cleavage in animal cells remain largely unexplored.

Purpose of the Study:

  • To investigate the in vivo occurrence of self-catalyzed cleavage in newt sat2 transcripts.
  • To determine if the hammerhead (HH) motif in sat2 RNA is functional within animal cells.
  • To characterize different sat2 transcript populations and their processing in various newt tissues.

Main Methods:

  • Analysis of synthetic and endogenous sat2 transcripts from various newt tissues (testes, liver, ovary).

Related Experiment Videos

  • Determination of transcript ends and end-group analysis to identify cleavage sites and products.
  • Comparison of in vivo transcript characteristics with in vitro self-cleavage products.

    • Main Results:

    • Two distinct sat2 transcript populations were identified.
    • Monomeric transcripts from testes and liver exhibit ends consistent with in vitro HH-mediated self-cleavage.
    • Ovarian transcripts, while not utilizing the canonical HH cleavage site, show evidence of processing.

    Conclusions:

    • This study provides the first evidence of hammerhead (HH) motif-mediated self-cleavage occurring in animal cells.
    • Sat2 transcripts in newts undergo in vivo processing, with evidence of both canonical HH cleavage and alternative processing pathways.
    • The findings suggest a conserved RNA self-cleavage mechanism across different kingdoms of life.