Fluorescence imaging reveals the nuclear behavior of peroxisome proliferator-activated receptor/retinoid X receptor heterodimers in the absence and presence of ligand.

Details

Serval ID
serval:BIB_312147D3381D
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Fluorescence imaging reveals the nuclear behavior of peroxisome proliferator-activated receptor/retinoid X receptor heterodimers in the absence and presence of ligand.
Journal
Journal of Biological Chemistry
Author(s)
Feige J.N., Gelman L., Tudor C., Engelborghs Y., Wahli W., Desvergne B.
ISSN
0021-9258
Publication state
Published
Issued date
2005
Peer-reviewed
Oui
Volume
280
Number
18
Pages
17880-17890
Language
english
Notes
Publication types: Journal Article
Abstract
In a global approach combining fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), and fluorescence resonance energy transfer (FRET), we address the behavior in living cells of the peroxisome proliferator-activated receptors (PPARs), a family of nuclear receptors involved in lipid and glucose metabolism, inflammation control, and wound healing. We first demonstrate that unlike several other nuclear receptors, PPARs do not form speckles upon ligand activation. The subnuclear structures that may be observed under some experimental conditions result from overexpression of the protein and our immunolabeling experiments suggest that these structures are subjected to degradation by the proteasome. Interestingly and in contrast to a general assumption, PPARs readily heterodimerize with retinoid X receptor (RXR) in the absence of ligand in living cells. PPAR diffusion coefficients indicate that all the receptors are engaged in complexes of very high molecular masses and/or interact with relatively immobile nuclear components. PPARs are not immobilized by ligand binding. However, they exhibit a ligand-induced reduction of mobility, probably due to enhanced interactions with cofactors and/or chromatin. Our study draws attention to the limitations and pitfalls of fluorescent chimera imaging and demonstrates the usefulness of the combination of FCS, FRAP, and FRET to assess the behavior of nuclear receptors and their mode of action in living cells.
Keywords
Active Transport, Cell Nucleus/physiology, Animals, COS Cells, Cell Nucleus/chemistry, Cell Nucleus/metabolism, Cercopithecus aethiops, Fluorescence Resonance Energy Transfer/methods, Humans, Ligands, Peroxisome Proliferator-Activated Receptors/analysis, Peroxisome Proliferator-Activated Receptors/metabolism, Retinoid X Receptors/analysis, Retinoid X Receptors/metabolism
Pubmed
Web of science
Open Access
Yes
Create date
24/01/2008 15:27
Last modification date
20/08/2019 13:16
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