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.

[1]  Ellson Y. Chen,et al.  Overview of manual and automated DNA sequencing by the dideoxy chain termination method , 1991 .

[2]  A. Glazer,et al.  Identification and Location of ε-N-Trimethyllysine in Yeast Cytochromes c , 1970 .

[3]  E. Phizicky,et al.  A 2′-Phosphotransferase Implicated in tRNA Splicing Is Essential in Saccharomyces cerevisiae * , 1997, The Journal of Biological Chemistry.

[4]  E. Phizicky,et al.  An enzyme from Saccharomyces cerevisiae uses NAD+ to transfer the splice junction 2'-phosphate from ligated tRNA to an acceptor molecule. , 1991, The Journal of biological chemistry.

[5]  J. Abelson,et al.  Mechanism of action of a yeast RNA ligase in tRNA splicing , 1983, Cell.

[6]  J. Abelson,et al.  Precise excision of intervening sequences from precursor tRNAs by a membrane-associated yeast endonuclease , 1983, Cell.

[7]  M. Meaney,et al.  Nongenomic transmission across generations of maternal behavior and stress responses in the rat. , 1999, Science.

[8]  E. Phizicky,et al.  Yeast tRNA ligase mutants are nonviable and accumulate tRNA splicing intermediates. , 1992, The Journal of biological chemistry.

[9]  F. Sherman,et al.  Yeast cytochrome c-specific protein-lysine methyltransferase: coordinate regulation with cytochrome c and activities in cyc mutants , 1979, Journal of bacteriology.

[10]  I. Macreadie,et al.  Improved shuttle vectors for cloning and high-level Cu(2+)-mediated expression of foreign genes in yeast. , 1991, Gene.

[11]  D. Turner,et al.  Transient ADP-ribosylation of a 2′-Phosphate Implicated in Its Removal from Ligated tRNA during Splicing in Yeast* , 1999, The Journal of Biological Chemistry.

[12]  H. Lodish,et al.  Cloning by function: expression cloning in mammalian cells. , 1994, Trends in pharmacological sciences.

[13]  F. Sherman Getting started with yeast. , 1991, Methods in enzymology.

[14]  T. Kawamoto,et al.  Purification and Properties of Calmodulin‐Lysine N‐Methyltransferase from Rat Brain Cytosol , 1987, Journal of neurochemistry.

[15]  R. Rauhut,et al.  The Yeast tRNA Splicing Endonuclease: A Tetrameric Enzyme with Two Active Site Subunits Homologous to the Archaeal tRNA Endonucleases , 1997, Cell.

[16]  B. Edgar,et al.  Developmental Control of Cell Cycle Regulators: A Fly's Perspective , 1996, Science.

[17]  J. Hudson,et al.  The complete set of predicted genes from Saccharomyces cerevisiae in a readily usable form. , 1997, Genome research.

[18]  C. Lawrence,et al.  Thymine-Thymine Dimer Bypass by Yeast DNA Polymerase ζ , 1996, Science.

[19]  E. Phizicky,et al.  A highly specific phosphatase from Saccharomyces cerevisiae implicated in tRNA splicing , 1990, Molecular and cellular biology.

[20]  E. Phizicky,et al.  A conditional lethal yeast phosphotransferase (tpt1) mutant accumulates tRNAs with a 2'-phosphate and an undermodified base at the splice junction. , 1997, RNA.

[21]  W. Filipowicz,et al.  tRNA splicing in yeast and wheat germ. A cyclic phosphodiesterase implicated in the metabolism of ADP-ribose 1",2"-cyclic phosphate. , 1994, The Journal of biological chemistry.

[22]  D. Turner,et al.  An NAD derivative produced during transfer RNA splicing: ADP-ribose 1"-2" cyclic phosphate. , 1993, Science.