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Vol. 13, Issue 6, 2001-2015, June 2002
Laboratory of Receptor Biology and Gene Expression, National Cancer
Institute, National Institutes of Health, Bethesda, Maryland 20892-5055
The aryl hydrocarbon receptor (AhR or dioxin receptor) is a
ligand-activated transcription factor that heterodimerizes with the AhR
nuclear translocator (ARNT/HIF-1
) to form an AhR/ARNT transcription
factor complex. This complex binds to specific DNA sites in the
regulatory domains of numerous target genes and mediates the biological
effects of exogenous ligands. Herein, we have investigated the
subcellular distribution of the AhR/ARNT complex in response to ligand
stimulation, by using live-cell confocal and high-resolution deconvolution microscopy. We found that unliganded AhR shows a predominantly cytoplasmic diffuse distribution in mouse hepatoma cells.
On addition of ligand, AhR rapidly translocates to the nucleus and
accumulates in multiple bright foci. Inhibition of transcription
prevented the formation of AhR foci. Dual- and triple-immunolabeling experiments, combined with labeling of nascent RNA, showed that the
foci are transcription sites, indicating that upon ligand stimulation,
AhR is recruited to active transcription sites. The interaction of AhR
with ARNT was both necessary and sufficient for the recruitment of AhR
to transcription sites. These results indicate that AhR/ARNT complexes
are recruited to specific subnuclear compartments in a ligand-dependent
manner and that these foci represent the sites of AhR target genes.
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