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Rapid Synthesis and Screening of Chemically Activated Transcription Factors with GFP-based Reporters
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NADP regulates the yeast GAL induction system.

P Rajesh Kumar1, Yao Yu, Rolf Sternglanz

  • 1Cold Spring Harbor Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY11724, USA.

Science (New York, N.Y.)
|February 23, 2008
PubMed
Summary

Nicotinamide adenine dinucleotide phosphate (NADP) is a key factor in regulating galactose metabolism genes in yeast. This dinucleotide mediates the interaction between Gal4p and Gal80p, explaining rapid gene induction.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Gene Regulation

Background:

  • Transcriptional regulation of galactose metabolism in Saccharomyces cerevisiae involves Gal4p, Gal80p, and Gal3p.
  • The existing model of induction via cytoplasmic sequestration of Gal80p by Gal3p does not fully explain the rapid response observed.
  • A missing factor in the current model of transcriptional regulation was suspected.

Purpose of the Study:

  • To elucidate the molecular mechanism underlying the rapid transcriptional induction of galactose-metabolizing genes in yeast.
  • To identify the missing factor involved in the interaction between Gal4p and Gal80p.
  • To characterize the role of any identified factors in the Gal4p-Gal80p regulatory complex.

Main Methods:

  • Structural analysis of the Gal80p-Gal4p interaction using X-ray crystallography.
  • Biochemical assays to investigate molecular interactions.
  • In vivo experiments in Saccharomyces cerevisiae to validate findings.

Main Results:

  • The crystal structure of Gal80p bound to a Gal4p peptide revealed an associated dinucleotide.
  • Biochemical and in vivo studies identified nicotinamide adenine dinucleotide phosphate (NADP) as this mediating dinucleotide.
  • NADP was shown to play a crucial role in the initial induction event of the galactose regulon.

Conclusions:

  • Nicotinamide adenine dinucleotide phosphate (NADP) is a critical component in the transcriptional regulation of galactose metabolism in yeast.
  • NADP mediates the interaction between the transcriptional activator Gal4p and the repressor Gal80p.
  • The discovery of NADP's role provides a more complete understanding of the rapid induction mechanism for galactose-inducible genes.