A biochemical genomics approach for identifying genes by the activity of their products.
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.
Martzen
M R
MR
Department of Biochemistry and Biophysics, University of Rochester School of Medicine, Rochester, NY 14642, USA.
McCraith
S M
SM
Spinelli
S L
SL
Torres
F M
FM
Fields
S
S
Grayhack
E J
EJ
Phizicky
E M
EM
D000246:Adenosine Diphosphate Ribose / Q000031:analogs & derivatives / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D005821:Genetic Techniques; D005982:Glutathione Transferase / Q000235:genetics; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016366:Open Reading Frames; D010727:Phosphoric Diester Hydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D012680:Sensitivity and Specificity
D005656:Fungal Proteins; D011993:Recombinant Fusion Proteins; C401161:adenosine diphosphate-ribose 1''-2''cyclic phosphate; D000246:Adenosine Diphosphate Ribose; C034411:cytochrome C methyltransferase; D011495:Histone-Lysine N-Methyltransferase; D005982:Glutathione Transferase; D010727:Phosphoric Diester Hydrolases