title: Chemical complementation: small-molecule-based genetic selection in yeast.
abstract: Protein and metabolic engineering would greatly benefit from a general system linking the presence of a small molecule to the power of genetic selection. We use nuclear receptors to link the survival of Saccharomyces cerevisiae to the presence of small molecules through genetic selection, extending classical genetic complementation to a new "chemical complementation." In this system the Gal4 DNA-binding domain is fused to ligand-binding domains from two nuclear receptors, expressed in the strain PJ69-4A, and grown on plates containing known ligands for the receptors. Yeast survive on selective plates only in the presence of a nuclear receptor and the corresponding ligand. Mutagenesis can increase the sensitivity of chemical complementation. This system may be extended to engineer nuclear receptors for practically any small molecule through directed evolution coupled to genetic selection, and for performing metabolic engineering in yeast.
authors.affiliation: School of Chemistry and Biochemistry, Georgia Institute of Technology, Atlanta, GA 30332, USA.
authors.forename: Bahareh
authors.initials: B
authors.lastname: Azizi
authors.forename: Eileen I
authors.initials: EI
authors.lastname: Chang
authors.forename: Donald F
authors.initials: DF
authors.lastname: Doyle
mesh_terms: D000970:Antineoplastic Agents / Q000378:metabolism; D000975:Antioxidants / Q000378:metabolism; D004268:DNA-Binding Proteins; D005816:Genetic Complementation Test; D008024:Ligands; D015394:Molecular Structure; D016297:Mutagenesis, Site-Directed; D017239:Paclitaxel / Q000378:metabolism; D000077297:Pregnane X Receptor; D011485:Protein Binding; D018160:Receptors, Cytoplasmic and Nuclear / Q000235:genetics / Q000378:metabolism; D018168:Receptors, Retinoic Acid / Q000235:genetics / Q000378:metabolism; D011987:Receptors, Steroid / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D047488:Retinoid X Receptors; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D012641:Selection, Genetic; D024505:Tocopherols / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*; D014212:Tretinoin / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D000975:Antioxidants; D004268:DNA-Binding Proteins; C052130:GAL4 protein, S cerevisiae; D008024:Ligands; D000077297:Pregnane X Receptor; D018160:Receptors, Cytoplasmic and Nuclear; D018168:Receptors, Retinoic Acid; D011987:Receptors, Steroid; D011993:Recombinant Fusion Proteins; D047488:Retinoid X Receptors; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D014212:Tretinoin; D017239:Paclitaxel; D024505:Tocopherols