Drosophila nonsense suppressors: functional analysis in Saccharomyces cerevisiae, Drosophila tissue culture cells and Drosophila melanogaster.
Amber (UAG) and opal (UGA) nonsense suppressors were constructed by oligonucleotide site-directed mutagenesis of two Drosophila melanogaster leucine-tRNA genes and tested in yeast, Drosophila tissue culture cells and transformed flies. Suppression of a variety of amber and opal alleles occurs in yeast. In Drosophila tissue culture cells, the mutant tRNAs suppress hsp70:Adh (alcohol dehydrogenase) amber and opal alleles as well as an hsp70:beta-gal (beta-galactosidase) amber allele. The mutant tRNAs were also introduced into the Drosophila genome by P element-mediated transformation. No measurable suppression was seen in histochemical assays for Adhn4 (amber), AdhnB (opal), or an amber allele of beta-galactosidase. Low levels of suppression (approximately 0.1-0.5% of wild type) were detected using an hsp70:cat (chloramphenicol acetyltransferase) amber mutation. Dominant male sterility was consistently associated with the presence of the amber suppressors.
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Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110-1095.
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D000426:Alcohol Dehydrogenase / Q000037:antagonists & inhibitors / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D004331:Drosophila melanogaster / Q000235:genetics*; D008969:Molecular Sequence Data; D009154:Mutation; D012356:RNA, Transfer, Leu / Q000235:genetics; D012098:Reproduction / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D013489:Suppression, Genetic; D014170:Transformation, Genetic
D004251:DNA Transposable Elements; D012356:RNA, Transfer, Leu; D000426:Alcohol Dehydrogenase