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Investigating Interactions Between Histone Modifying Enzymes and Transcription Factors in vivo by Fluorescence Resonance Energy Transfer
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Investigating Interactions Between Histone Modifying Enzymes and Transcription Factors in vivo by Fluorescence Resonance Energy Transfer

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Imaging Erg and Jun transcription factor interaction in living cells using fluorescence resonance energy transfer

Barbara Camuzeaux1, Corentin Spriet, Laurent Héliot

  • 1UMR 8526CNRS/Institut Pasteur de Lille/Université de Lille2, Institut de Biologie de Lille, BP 447, 1 rue Calmette, 59021 Lille cedex, France.

Biochemical and Biophysical Research Communications
|June 1, 2005
PubMed

Insights

Transcription factors Erg and Jun interact in living cells, a key step for their synergistic gene regulation. This interaction depends on a specific residue (Y371) in the Erg protein

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Genetics

Background:

  • Transcription factors regulate gene expression through physical interactions.
  • Previous in vitro studies suggested synergy between Erg (Ets family) and Jun/Fos (bZip family).
  • Direct Erg-Jun protein interactions are crucial for this synergy.

Purpose of the Study:

  • To visualize and confirm protein-protein interactions between Erg and Jun in living cells.
  • To investigate the role of specific protein domains and residues in mediating these interactions.
  • To establish a link between protein interaction dynamics and transcriptional synergy.

Main Methods:

  • Generation of fusion proteins: Erg, Fos, and Jun tagged with yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP).
  • Transient expression in HeLa cells.
  • Fluorescence Resonance Energy Transfer (FRET) microscopy, including photobleaching FRET and fluorescence lifetime imaging microscopy (FLIM), in fixed and living cells.

Main Results:

  • Demonstrated intermolecular FRET between YFP-Erg and CFP-Jun in living cells.
  • Confirmed Erg-Jun interaction using two distinct FRET techniques.
  • Showed a loss of FRET signal when using an Erg mutant (Y371F) in the Ets domain, indicating this residue's importance.
  • Provided evidence for the direct interaction of Erg and Jun proteins in a cellular context.

Conclusions:

  • Erg and Jun proteins directly interact within living cells.
  • This interaction is a prerequisite for their observed transcriptional synergy.
  • The Y371 residue in the Erg protein's Ets domain is essential for mediating the Erg-Jun interaction.

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