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Related Experiment Videos

GAL4 is regulated by a glucose-responsive functional domain

G Stone1, I Sadowski

  • 1Department of Biochemistry, University of British Columbia, Vancouver, Canada.

The EMBO Journal
|April 1, 1993
PubMed
Summary

Glucose directly inhibits the Saccharomyces cerevisiae GAL4 protein via its central region. This mechanism, involving inhibitory and glucose response domains, rapidly switches gene activation from galactose to glucose utilization.

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Area of Science:

  • Molecular Biology
  • Yeast Genetics
  • Gene Regulation

Background:

  • The GAL4 protein activates transcription in Saccharomyces cerevisiae.
  • GAL4 activity is modulated by carbon sources like glucose and galactose.
  • Glucose, the preferred carbon source, inhibits GAL4 function through various mechanisms.

Purpose of the Study:

  • To investigate the direct inhibitory mechanisms of glucose on GAL4 activity.
  • To identify specific regions within GAL4 responsible for glucose-mediated repression.
  • To elucidate the role of these regions in the carbon source switch.

Main Methods:

  • Deletion analysis of the GAL4 central region.
  • Fusion of GAL4 central region to a heterologous activator.
  • Assays for direct inhibition of GAL4 activity upon glucose addition.

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Main Results:

  • A previously uncharacterized central region of GAL4 contains inhibitory domains and a glucose response domain (GRD).
  • Deletion of this central region abolished glucose-mediated direct inhibition of GAL4.
  • The central region conferred glucose-dependent inhibition when fused to another activator.
  • Constitutive inhibition was observed when the GRD was absent.

Conclusions:

  • Glucose directly inhibits GAL4 activity through its central region.
  • This direct inhibition is rapid, occurring within 30 minutes of glucose addition.
  • The central region, including inhibitory domains and the GRD, plays a key role in the switch to glucose utilization.
  • A model involving interaction with another protein is proposed for glucose-mediated GAL4 repression.