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Submitted on October 30, 2002
Accepted on February 21, 2003
1 Research Institute, Saitama Cancer Center, 818 Komuro, Ina-machi, Kita-adachi, Saitama 362-0806, JAPAN; Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation, Kawaguchi 332-0012, JAPAN; Department of Developmental Biology, National Institute for Basic Biology, Myodaiji-cho, Okazaki 444-8585, JAPAN; Department of Molecular Biomechanics, School of Life Science, The Graduate University for Advanced Studies, Myodaiji-cho, Okazaki 444-8585, JAPAN; Department of Biochemistry, Saitama Medical School, 38 Morohongo, Moroyama-machi, Iruma-gun, Saitama 350-0451, JAPAN; Department of Pediatrics, Asahikawa Medical College, Midorigaoka, Higashi, Asahikawa 078-8510, JAPAN.
* To whom correspondence should be addressed. E-mail: moro{at}nibb.ac.jp.
Dax-1 (NR0B1) is an orphan nuclear receptor that represses transcription by Ad4BP/SF-1 (NR5A1). Observations on human diseases and the phenotypes of mice, in which the corresponding genes have been disrupted, have elucidated essential roles of these two nuclear receptors in steroidogenic tissues differentiation. However, little is known about how the functions of these factors are regulated. Here we have examined their subcellular localization, and have clarified the molecular mechanisms regulating subcellular localization of Dax-1. Prompted by the finding that nuclear localization of Dax-1 correlates with the presence of Ad4BP/SF-1 in the early stages of pituitary development, we have tested the possibility that interaction between the two factors is essential for the nuclear localization of Dax-1. In vitro studies with cultured cells demonstrated that an interaction involving the LXXLL motifs in the N-terminal repeat region of Dax-1 plays a key role in its subcellular localization. In addition, we found that a mutant form of DAX-1, [L466R], from a patient with adrenal hypoplasia congenita, was defective in nuclear localization in spite of having an intact N-terminus. Taken together the results reveal that the subcellular localization of Dax-1 is influenced by the presence of Ad4BP/SF-1, and that two regions of Dax-1 have important roles for this process.
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