title: A genome-wide overexpression screen in yeast for small-molecule target identification.
abstract: We describe a multicopy gene suppression screen of drug sensitivity in Saccharomyces cerevisiae that facilitates the identification of cellular targets of small molecules. An array of yeast transformants harboring a multicopy yeast genomic library was screened for resistance to growth inhibitors. Comparison of array growth patterns for several such inhibitors allowed the differentiation of general and molecule-specific genetic suppressors. Specific resistance to phenylaminopyrimidine (1), an inhibitor identified from a kinase-directed library, was associated with the overexpression of Pkc1 and a subset of downstream kinases. Components of two other pathways (pheromone response/filamentous growth and Pho85 kinase) that genetically interact with the PKC1 MAPK signaling cascade were also identified. Consistent with the suppression screen, inhibitor 1 bound to Pkc1 in yeast cell lysate and inhibited its activity in vitro. These results demonstrate the utility of this approach for the rapid deconvolution of small-molecule targets.
authors.affiliation: Department of Chemistry, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA.
authors.forename: Hendrik
authors.initials: H
authors.lastname: Luesch
authors.forename: Tom Y H
authors.initials: TY
authors.lastname: Wu
authors.forename: Pingda
authors.initials: P
authors.lastname: Ren
authors.forename: Nathanael S
authors.initials: NS
authors.lastname: Gray
authors.forename: Peter G
authors.initials: PG
authors.lastname: Schultz
authors.forename: Frantisek
authors.initials: F
authors.lastname: Supek
mesh_terms: D004353:Drug Evaluation, Preclinical / Q000379:methods; D025141:Drug Resistance, Fungal / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000187:drug effects; D015698:Genomic Library; D020935:MAP Kinase Signaling System / Q000187:drug effects / Q000502:physiology; D008826:Microbial Sensitivity Tests; D015394:Molecular Structure; D008970:Molecular Weight; D011493:Protein Kinase C / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D011725:Pyridines / Q000138:chemical synthesis / Q000494:pharmacology*; D011743:Pyrimidines / Q000138:chemical synthesis / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013012:Sorbitol / Q000494:pharmacology; D013997:Time Factors
chemical_list: C502731:N-(3-chlorophenyl)-4-(4-pyridinyl)-2-pyrimidinamine; D011725:Pyridines; D011743:Pyrimidines; D029701:Saccharomyces cerevisiae Proteins; D013012:Sorbitol; C490702:PKC1 protein, S cerevisiae; D011493:Protein Kinase C