Imaging Erg and Jun transcription factor interaction in living cells using fluorescence resonance energy transfer analyses
- UMR 8526CNRS/Institut Pasteur de Lille/Universite de Lille2, Institut de Biologie de Lille, BP 447, 1 rue Calmette, 59021 Lille cedex (France)
- Service d'Imagerie Cellulaire Fonctionnelle, FRC3 CNRS, Institut de Biologie de Lille, BP 447, 1 rue Calmette, 59021 Lille cedex (France)
- UMR 8527CNRS/Institut Pasteur de Lille/Universite de Lille2, Institut de Biologie de Lille, BP 447, 1 rue Calmette, 59021 Lille cedex (France)
Physical interactions between transcription factors play important roles in modulating gene expression. Previous in vitro studies have shown a transcriptional synergy between Erg protein, an Ets family member, and Jun/Fos heterodimer, members of the bZip family, which requires direct Erg-Jun protein interactions. Visualization of protein interactions in living cells is a new challenge in biology. For this purpose, we generated fusion proteins of Erg, Fos, and Jun with yellow and cyan fluorescent proteins, YFP and CFP, respectively. After transient expression in HeLa cells, interactions of the resulting fusion proteins were explored by fluorescence resonance energy transfer microscopy (FRET) in fixed and living cells. FRET between YFP-Erg and CFP-Jun was monitored by using photobleaching FRET and fluorescence lifetime imaging microscopy. Both techniques revealed the occurrence of intermolecular FRET between YFP-Erg and CFP-Jun. This is stressed by loss of FRET with an YFP-Erg version carrying a point mutation in its ETS domain. These results provide evidence for the interaction of Erg and Jun proteins in living cells as a critical prerequisite of their transcriptional synergy, but also for the essential role of the Y371 residue, conserved in most Ets proteins, in this interaction.
- OSTI ID:
- 20710852
- Journal Information:
- Biochemical and Biophysical Research Communications, Vol. 332, Issue 4; Other Information: DOI: 10.1016/j.bbrc.2005.05.057; PII: S0006-291X(05)00983-6; Copyright (c) 2005 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA); ISSN 0006-291X
- Country of Publication:
- United States
- Language:
- English
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