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Related Experiment Videos

GAGA factor down-regulates its own promoter.

Ana Kosoy1, Sara Pagans, Maria Lluisa Espinas

  • 1Departament de Biologia Molecular i Cel.lular, Institut de Biologia Molecular de Barcelona, Consell Superior d'Investigacions Cientifiques, Jordi Girona, 18-26, Spain.

The Journal of Biological Chemistry
|August 30, 2002
PubMed
Summary

GAGA factor, a known transcription activator, surprisingly represses its own Trl gene expression in Drosophila. This DNA-binding dependent repression was confirmed using RNA interference in S2 cells.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Developmental Biology

Background:

  • GAGA factor is a known transcription activator in Drosophila.
  • Its role in regulating its own gene (Trl) promoter is not well understood.

Purpose of the Study:

  • To investigate the regulatory role of GAGA factor on the Trl promoter.
  • To determine if GAGA factor can repress Trl gene expression.

Main Methods:

  • Transient transfection assays in S2 cells to define the minimal Trl promoter.
  • DNase I footprinting to identify GAGA binding sites.
  • Chromatin immunoprecipitation (ChIP) to confirm GAGA binding in vivo.
  • Double-stranded RNA interference (dsRNAi) to assess endogenous GAGA function.

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Main Results:

  • A minimal Trl promoter was identified with multiple GAGA binding sites.
  • GAGA factor was found to down-regulate Trl expression in a DNA-binding dependent manner.
  • GAGA factor binds to the Trl promoter region in S2 cells.
  • Endogenous GAGA factors limit Trl expression.

Conclusions:

  • GAGA factor acts as a repressor of its own Trl gene expression.
  • This repression mechanism is dependent on GAGA factor binding to DNA.
  • The findings suggest GAGA factor may repress other promoters as well.