title: Light-mediated control of DNA transcription in yeast.
abstract: A variety of methods exist for inducible control of DNA transcription in yeast. These include the use of native yeast promoters or regulatory elements that are responsive to small molecules such as galactose, methionine, and copper, or engineered systems that allow regulation by orthogonal small molecules such as estrogen. While chemically regulated systems are easy to use and can yield high levels of protein expression, they often provide imprecise control over protein levels. Moreover, chemically regulated systems can affect many other proteins and pathways in yeast, activating signaling pathways or physiological responses. Here, we describe several methods for light mediated control of DNA transcription in vivo in yeast. We describe methodology for using a red light and phytochrome dependent system to induce transcription of genes under GAL1 promoter control, as well as blue light/cryptochrome dependent systems to control transcription of genes under GAL1 promoter or LexA operator control. Light is dose dependent, inexpensive to apply, easily delivered, and does not interfere with cellular pathways, and thus has significant advantages over chemical systems.
authors.affiliation: Department of Biology, Duke University, Durham, NC, United States.
authors.forename: Robert M
authors.initials: RM
authors.lastname: Hughes
authors.forename: Steven
authors.initials: S
authors.lastname: Bolger
authors.forename: Hersh
authors.initials: H
authors.lastname: Tapadia
authors.forename: Chandra L
authors.initials: CL
authors.lastname: Tucker
mesh_terms: D029681:Arabidopsis Proteins / Q000096:biosynthesis / Q000235:genetics; D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis / Q000235:genetics; D056931:Cryptochromes / Q000096:biosynthesis / Q000235:genetics; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D005686:Galactokinase / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000528:radiation effects*; D017930:Genes, Reporter; D008027:Light; D052980:Phycobilins / Q000494:pharmacology / Q000528:radiation effects; D010798:Phycocyanin / Q000494:pharmacology / Q000528:radiation effects; D051939:Phytochrome B / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis / Q000235:genetics; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic / Q000528:radiation effects*; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
chemical_list: D029681:Arabidopsis Proteins; D051792:Basic Helix-Loop-Helix Proteins; D056931:Cryptochromes; D004268:DNA-Binding Proteins; D005686:Galactokinase; D052980:Phycobilins; D010798:Phycocyanin; D051939:Phytochrome B; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; D001616:beta-Galactosidase; C533832:CIB1 protein, Arabidopsis; C103618:CRY2 protein, Arabidopsis; C052130:GAL4 protein, S cerevisiae; C497952:PHYB protein, Arabidopsis; C116162:PIF3 protein, Arabidopsis; C011069:phycocyanobilin; C524219:GAL1 protein, S cerevisiae