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Am J Physiol Endocrinol Metab (January 21, 2004). doi:10.1152/ajpendo.00508.2003
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Submitted on November 11, 2003
Accepted on January 20, 2004

Mitochondrial Localization of ER{alpha} and ER{beta} in Human MCF-7 cells

Jin Q. Chen1*, Michael Delannoy2, Carol Cooke2, and James D. Yager1

1 Department of Environmental Health Sciences, Division of Toxicological Sciences, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA
2 Microscopy Facility, Johns Hopkins School of Medicine, Baltimore, MD, USA

* To whom correspondence should be addressed. E-mail: jichen{at}jhsph.edu.

We observed previously that estrogen treatment increased the transcript levels of several mitochondrial DNA (mtDNA) encoded genes for mitochondrial respiratory chain (MRC) proteins and MRC activity in rat hepatocytes and human HepG2 cells. Others have reported detection of estrogen receptors (ER), ER{alpha} and ER{beta}, in mitochondria of rabbit ovarian and uterine tissue. In this study, we have extended these observations. Using cellular fractionation and Western blot with ER{alpha}- and ER{beta}-specific antibodies, we observed that ER{alpha} and ER{beta} are present in mitochondria of human MCF-7 cells and that the mitochondrial ER{alpha} and ER{beta} account for 10% and 18%, respectively, of total cellular ER{alpha} and ER{beta} in E2-treated MCF-7 cells. We also found that E2 significantly enhanced the amounts of mitochondrial ER{alpha} and ER{beta} in a time- and concentration-dependent manner and that these effects are accompanied by a significant increase in the transcript levels of mtDNA-encoded genes, i.e. cytochrome c oxidase subunits I and II. Moreover, we demonstrated that these E2-mediated effects were inhibited by the pure ER antagonist, ICI182780, indicating the involvement of ERs. Using immunohistochemistry with confocal microscopy and immunogold electron microscopy, we demonstrated that ER{alpha} and ER{beta} are located within the MCF-7 cell mitochondrial matrix. Computer analysis identified a putative internal mitochondrial targeting peptide signal within human ER{beta}, suggesting an inherent potential for ER{beta} to enter mitochondria. These findings confirm the observations of others and provide additional support for this novel localization of the ERs and for a potentially important role of the ER in the regulation of mtDNA transcription.




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