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J. Biol. Chem., Vol. 277, Issue 42, 39144-39155, October 18, 2002
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From the The human P450scc gene is regulated by the
tissue-specific orphan nuclear receptor, steroidogenic factor-1 (SF-1),
which plays a key role in several physiologic processes including
steroid synthesis, adrenal and gonadal development, and sexual
differentiation. Several studies have demonstrated the interaction of
SF-1 with different proteins. However, it is clear that additional
factors not yet identified are involved with SF-1 to regulate different target genes. Recently, it was demonstrated that a novel
transcriptional regulating protein of 132 kDa (TReP-132) regulates
expression of the human P450scc gene. The overexpression of TReP-132 in
adrenal cells increases the production of pregnenolone, which is
associated with the activation of P450scc gene expression. Considering
the colocalization of TReP-132 and SF-1 in steroidogenic tissues such as the adrenal and testis, and the presence of two putative
LXXLL motifs in TReP-132 that can potentially
interact with SF-1, the relationship between these two factors on the
P450scc gene promoter was determined. The coexpression of SF-1 and
TReP-132 in adrenal NCI-H295 cells cooperates to increase promoter
activity. Pull-down experiments demonstrated the interaction between
TReP-132 and SF-1, and this was further confirmed in intact cells by
coimmunoprecipitation/Western blot and two-hybrid analyses. Deletions
and mutations of the TReP-132 cDNA sequence demonstrate that SF-1
interaction requires the LXXLL motif found at the
amino-terminal region of the protein. Also, the "proximal activation
domain" and the "AF-2 hexamer" motif of SF-1 are involved in
interaction with TReP-132. Consistent with previous studies showing
interaction between CBP/p300 and SF-1 or TReP-132, the coexpression of
these three proteins results in a synergistic effect on P450scc gene
promoter activity. Taken together the results in this study
identify a novel function of TReP-132 as a partner in a complex with
SF-1 and CBP/p300 to regulate gene transcription involved in steroidogenesis.
The Transcriptional Regulating Protein of 132 kDa
(TReP-132) Enhances P450scc Gene Transcription through Interaction
with Steroidogenic Factor-1 in Human Adrenal Cells*
,,,¶
Oncology and Molecular Endocrinology
Research Center, Laval University, Québec GIK 7P4,
Canada, § U545 INSERM, Département
d'Athérosclérose, Institut Pasteur de Lille, and
Faculté de Pharmacie, Université de Lille II,
Lille 59019, France, and ¶ Genfit, Parc Eurasanté,
59120 Loos, France
*
This work was supported in part by Operating Grant MT-12901
from the Medical Research Council of Canada and by Scholarship 980149 from the Fonds de la Recherche en Santé du Québec (FRSQ) awarded to D. W. H.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Genfit, Parc
Eurasanté, 885, Ave. Eugène Avinée, 59120 Loos,
France. Tel.: 33-3-20-16-40-00; Fax: 33-3-20-16-40-01; E-mail:
Dean.Hum@genfit.com.
Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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