LXXLL-related motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1

Taiga Suzuki, Megumi Kasahara, Hidefumi Yoshioka, Ken ichirou Morohashi, Kazuhiko Umesono

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Abstract

The orphan receptor Ad4BP/SF-1 (NR5A1) is a constitutive activator, and its activity is repressed by another orphan receptor, Dax-1 (NR0B1). In the present study, we investigated the molecular mechanisms underlying this repression by Dax-1. Yeast two-hybrid and transient-transfection assays confirmed the necessity of three LXXLL-related motifs in Dax-1 for interaction with and repression of Ad4BP/SF-1. In vitro pull-down experiments confirmed that Dax-1 interacts with Ad4BP/SF-1 and also with LRH-1 (NR5A2). The target specificity of the LXXLL-related motifs was indicated by the observations that Ad4BP/SF-1, ERα (NR3A1), LRH-1, ERR2 (NR3B2), and fly FTZ-F1 (NR5A3) interacted through their ligand binding domains with all the LXXLL-related motifs in Dax-1 whereas HNF4 (NR2A1) and RORα (NR1F1) did not. Transcriptional activities of the receptors whose DNA binding domains (DBDs) were replaced by the GAL4 DBD were repressed by Dax-1 to various levels, which correlated with the strength of interaction. Amino acid substitutions revealed that Ad4BP/SF-1 and LRH-1 preferentially interact with L(+1)XXLL-related motifs containing serine, tyrosine, serine, and threonine at positions -2, +2, +3, and +6, respectively. Taken together, our results indicate that the specificities of LXXLL-related motifs in Dax-1 based on their amino acid sequences play an important role in regulation of orphan receptors.

Original languageEnglish
Pages (from-to)238-249
Number of pages12
JournalMolecular and cellular biology
Volume23
Issue number1
DOIs
Publication statusPublished - Jan 1 2003

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Orphan Nuclear Receptors
Serine
Threonine
Amino Acid Substitution
Transfection
Tyrosine
Amino Acid Sequence
Yeasts
Ligands
DNA
DNA receptor
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

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LXXLL-related motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1. / Suzuki, Taiga; Kasahara, Megumi; Yoshioka, Hidefumi; Morohashi, Ken ichirou; Umesono, Kazuhiko.

In: Molecular and cellular biology, Vol. 23, No. 1, 01.01.2003, p. 238-249.

Research output: Contribution to journalArticle

Suzuki, Taiga ; Kasahara, Megumi ; Yoshioka, Hidefumi ; Morohashi, Ken ichirou ; Umesono, Kazuhiko. / LXXLL-related motifs in Dax-1 have target specificity for the orphan nuclear receptors Ad4BP/SF-1 and LRH-1. In: Molecular and cellular biology. 2003 ; Vol. 23, No. 1. pp. 238-249.
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