The functional analysis of nonsense suppressors derived from in vitro engineered Saccharomyces cerevisiae tRNA Trp genes

A. L. Atkin, M. A. Riazi, C. L. Greer, K. L. Roy, J. B. Bell

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

Nonsense suppressors derived from Saccharomyces cerevisiae tRNATrp genes have not been identified by classical genetic screens, although one can construct efficient amber (am) suppressors from them by making the appropriate anticodon mutation in vitro. Herein, a series of in vitro constructed putative suppressor genes was produced to test if pre-tRNATrp processing difficulties could help to explain the lack of classical tRNATrp-based suppressors. It is clear that inefficient processing of introns from precursor tRNATrp, or inaccurate overall processing, may explain why some of these constructs fail to promote nonsense suppression in vivo. However, deficient processing must be only one of the reasons why classical tRNATrp-based suppressors have not been characterized, as suppression may still be extremely weak or absent in instances where the in vitro construct can lead to an accumulation of mature tRNATrp. Furthermore, suppression is also very weak in strains transformed with an intronless derivative of a putative tRNATrp ochre (oc) suppressor gene, wherein intron removal cannot pose a problem.

Original languageEnglish (US)
Pages (from-to)57-65
Number of pages9
JournalGene
Volume134
Issue number1
DOIs
StatePublished - Nov 30 1993

Keywords

  • Processing
  • in vivo
  • intron
  • mutagenesis
  • ochre
  • recombinant DNA
  • Ψ

ASJC Scopus subject areas

  • Genetics

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