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Submitted on March 19, 2002
Accepted on June 24, 2002
and Estrogen Receptor ß
1 Department of Anatomy and Neurobiology, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Kamigyo-ku, Kyoto 602-8566, Japan
* To whom correspondence should be addressed. E-mail: matsuken{at}basic.kpu-m.ac.jp.
To investigate the relationships between the loci expressing functions of estrogen receptor (ER) and that of ERß, we analyzed the subnuclear distribution of ER and ERß in response to ligand in single living cells using fusion proteins labeled with different spectral variants of green fluorescent protein. Upon activation with ligand treatment, fluorescent protein tagged (FP)-ERß redistributed from a diffuse to discrete pattern within the nucleus showing a similar time course as FP-ER, and colocalized with FP-ER in the same discrete cluster. Analysis using deletion mutants of ER suggested that the ligand-dependent redistribution of ER might occur through a large part of the receptor including at least the latter part of activation function (AF)-1, the DNA binding domain, nuclear matrix binding domain and AF-2/ligand binding domain. In addition, a single AF-1 region within ER homodimer, or a single DNA binding domain as well as AF-1 region within the ER/ERß heterodimer could be sufficient for the cluster formation. More than half of the discrete clusters of FP-ER and FP-ERß were colocalized with hyperacetylated histone H4 and a component of the chromatin remodeling complex, Brg-1, indicating that ERs clusters might be involved in structural changes of chromatin.
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