Light-mediated control of DNA transcription in yeast

Methods. 2012 Dec;58(4):385-91. doi: 10.1016/j.ymeth.2012.08.004. Epub 2012 Aug 15.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Arabidopsis Proteins / biosynthesis
  • Arabidopsis Proteins / genetics
  • Basic Helix-Loop-Helix Transcription Factors / biosynthesis
  • Basic Helix-Loop-Helix Transcription Factors / genetics
  • Cryptochromes / biosynthesis
  • Cryptochromes / genetics
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics
  • Galactokinase / genetics
  • Gene Expression Regulation, Fungal / radiation effects*
  • Genes, Reporter
  • Light*
  • Phycobilins / pharmacology
  • Phycobilins / radiation effects
  • Phycocyanin / pharmacology
  • Phycocyanin / radiation effects
  • Phytochrome B / biosynthesis
  • Phytochrome B / genetics
  • Promoter Regions, Genetic
  • Protein Binding
  • Protein Interaction Mapping / methods
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism
  • Saccharomyces cerevisiae / radiation effects
  • Saccharomyces cerevisiae Proteins / biosynthesis
  • Saccharomyces cerevisiae Proteins / genetics
  • Transcription Factors / biosynthesis
  • Transcription Factors / genetics
  • Transcription, Genetic / radiation effects*
  • Two-Hybrid System Techniques
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics

Substances

  • Arabidopsis Proteins
  • Basic Helix-Loop-Helix Transcription Factors
  • CIB1 protein, Arabidopsis
  • CRY2 protein, Arabidopsis
  • Cryptochromes
  • DNA-Binding Proteins
  • GAL4 protein, S cerevisiae
  • PHYB protein, Arabidopsis
  • PIF3 protein, Arabidopsis
  • Phycobilins
  • Recombinant Fusion Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Phycocyanin
  • Phytochrome B
  • phycocyanobilin
  • GAL1 protein, S cerevisiae
  • Galactokinase
  • beta-Galactosidase