Abstract
For the identification of yeast genes specifying biochemical activities, a genomic strategy that is rapid, sensitive, and widely applicable was developed with an array of 6144 individual yeast strains, each containing a different yeast open reading frame (ORF) fused to glutathione S-transferase (GST). For the identification of ORF-associated activities, strains were grown in defined pools, and GST-ORFs were purified. Then, pools were assayed for activities, and active pools were deconvoluted to identify the source strains. Three previously unknown ORF-associated activities were identified with this strategy: a cyclic phosphodiesterase that acts on adenosine diphosphate-ribose 1"-2" cyclic phosphate (Appr>p), an Appr-1"-p-processing activity, and a cytochrome c methyltransferase.
Publication types
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Adenosine Diphosphate Ribose / analogs & derivatives
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Adenosine Diphosphate Ribose / metabolism
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Fungal Proteins / genetics*
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Fungal Proteins / metabolism
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Genes, Fungal*
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Genetic Techniques*
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Glutathione Transferase / genetics
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Histone-Lysine N-Methyltransferase / genetics
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Histone-Lysine N-Methyltransferase / isolation & purification
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Histone-Lysine N-Methyltransferase / metabolism
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Open Reading Frames*
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Phosphoric Diester Hydrolases / genetics
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Phosphoric Diester Hydrolases / isolation & purification
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Phosphoric Diester Hydrolases / metabolism
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Recombinant Fusion Proteins / isolation & purification
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Saccharomyces cerevisiae / genetics*
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Sensitivity and Specificity
Substances
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Fungal Proteins
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Recombinant Fusion Proteins
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adenosine diphosphate-ribose 1''-2''cyclic phosphate
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Adenosine Diphosphate Ribose
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cytochrome C methyltransferase
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Histone-Lysine N-Methyltransferase
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Glutathione Transferase
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Phosphoric Diester Hydrolases