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A gender-specific mRNA encoding a cytotoxic ribonuclease contains a 3' UTR of unusual length and structure

S Chen1, S Y Le, D L Newton

  • 1Laboratory of Experimental and Computational Biology, Division of Basic Science, SAIC Frederick, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21702, USA.

Nucleic Acids Research
|June 28, 2000
PubMed

Insights

Researchers identified a novel cytotoxic ribonuclease (rapLR1) in frogs, related to the antitumor protein onconase. Unusual structures in its 3' untranslated region may regulate gene expression.

Area of Science:

  • Molecular Biology
  • Genetics
  • Biochemistry

Background:

  • Onconase is an antitumor protein found in Rana pipiens.
  • Cytotoxic ribonucleases play roles in cellular processes and have therapeutic potential.
  • The regulation of gene expression often involves untranslated regions of mRNA.

Purpose of the Study:

  • To clone and characterize a novel ribonuclease (rapLR1) from Rana pipiens.
  • To investigate the structural and functional significance of the 3' untranslated region (UTR) of rapLR1 mRNA.
  • To explore the potential regulatory mechanisms of rapLR1 expression.

Main Methods:

  • cDNA library construction and cloning from Rana pipiens liver.
  • Secondary structure analysis and prediction of RNA folding.
  • In vitro translation assays.
  • Western blot analysis for protein detection.

Main Results:

  • A cDNA encoding rapLR1, related to onconase, was isolated.
  • The rapLR1 cDNA 3' UTR contained highly stable, unusual stem-loop structures (unusual folding regions).
  • The rapLR1 transcript was specific to female frog liver mRNA, while onconase protein was found in oocytes.
  • Deletion of the 3' UTR enhanced in vitro translation of rapLR1.

Conclusions:

  • The unusual folding regions in the rapLR1 3' UTR likely play a role in regulating mRNA metabolism and/or translation.
  • RapLR1 represents a novel cytotoxic ribonuclease with potential implications in gene regulation.
  • Differential expression of rapLR1 transcript and onconase protein suggests distinct biological roles.

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