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Nonsense suppression in Schizosaccharomyces pombe: The S. pombe Sup3-e tRNA UGASer gene is active in S. cerevisiae

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Summary

The gene encoding the efficient UGA suppressor sup3-e of Schizosaccharomyces pombe was isolated by in vivo transformation of Saccharomyces cerevisiae UGA mutants with S. pombe sup3-e DNA. DNA from a clone bank of EcoRI fragments from a S. pombe sup3-e strain in the hybrid yeast vector YRp17 was used to transform the S. cerevisiae multiple auxotroph his4-260 leu2-2 trp1-1 to prototrophy. Transformants were isolated at a low frequency; they lost the ability to grow in minimal medium after passaging in non-selective media. This suggested the presence of the suppressor gene on the non-integrative plasmid. Plasmid DNA, isolated from the transformed S. cerevisiae cells and subsequently amplified in E. coli, transformed S. cerevisiae his4-260 leu2-2 trp1-1 to prototrophy. In this way a 2.4 kb S. pombe DNA fragment carrying the sup3-e gene was isolated. Sequence analysis revealed the presence of two tRNA coding regions separated by a spacer of only seven nucleotides. The sup3-e tRNA UGASer tRNA gene is followed by a sequence coding for the initiator tRNAMet. The transformation results demonstrate that the cloned S. pombe UGA suppressor is active in S. cerevisiae UGA mutant strains.

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Authors and Affiliations

  1. Molecular Biophysics and Biochemistry, Yale University, 06511, New Haven, Connecticut, USA

    Herbert Hottinger, David Pearson, Fumiaki Yamao, Vera Gamulin, Lynn Colley & Dieter Söll

  2. Department of Biological Sciences, University of Pittsburgh, 15260, Pittsburgh, Pennsylvania, USA

    Terrance Cooper

Authors
  1. Herbert Hottinger
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  2. David Pearson
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  3. Fumiaki Yamao
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  4. Vera Gamulin
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  5. Lynn Colley
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  6. Terrance Cooper
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  7. Dieter Söll
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Communicated by G.R. Fink

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Hottinger, H., Pearson, D., Yamao, F. et al. Nonsense suppression in Schizosaccharomyces pombe: The S. pombe Sup3-e tRNA UGASer gene is active in S. cerevisiae . Mol Gen Genet 188, 219–224 (1982). https://doi.org/10.1007/BF00332678

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  • DOI: https://doi.org/10.1007/BF00332678

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