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J Biol Chem, Vol. 275, Issue 4, 2852-2858, January 28, 2000
From the Department of Pediatrics and the Metabolic Research Unit,
University of California, San Francisco, San Francisco, California
94143-0978
The cholesterol side-chain cleavage enzyme,
cytochrome P450scc, initiates the biosynthesis of all steroid hormones.
Adrenal and gonadal strategies for P450scc gene transcription are
essentially identical and depend on the orphan nuclear receptor
steroidogenic factor-1, but the placental strategy for transcription of
P450scc employs cis-acting elements different from those used in the
adrenal strategy and is independent of steroidogenic factor-1. Because placental expression of P450scc is required for human pregnancy, we
sought factors that bind to the
Cloning of Factors Related to HIV-inducible LBP Proteins That
Regulate Steroidogenic Factor-1-independent Human Placental
Transcription of the Cholesterol Side-chain Cleavage Enzyme,
P450scc*
155/131 region of the human P450scc
promoter, which participates in its placental but not adrenal or
gonadal transcription. A yeast one-hybrid screen of 2.4 × 106 cDNA clones from human placental JEG-3 cells
yielded two unique clones; one is the previously described
transcription factor LBP-1b, which is induced by HIV, type I infection
of lymphocytes, and the other is a new factor, termed LBP-9, that
shares 83% amino acid sequence identity with LBP-1b. When expressed in
transfected yeast, both factors bound specifically to the 155/131
DNA; antisera to LBP proteins supershifted the LBP-9·DNA complex and
inhibited formation of the LBP-1b·DNA complex. Reverse
transcriptase-polymerase chain reaction detected LBP-1b in human
placental JEG-3, adrenal NCI-H295A, liver HepG2, cervical HeLa, and
monkey kidney COS-1 cells, but LBP-9 was detected only in JEG-3 cells.
When the 155/131 fragment was linked to a minimal promoter,
co-expression of LBP-1b increased transcription 21-fold in a
dose-dependent fashion, but addition of LBP-9 suppressed
the stimulatory effect of LBP-1b. The roles of LBP transcription
factors in normal human physiology have been unclear. Their
modulation of placental but not adrenal P450scc transcription
underscores the distinctiveness of placental strategies for
steroidogenic enzyme gene transcription.
*
This work was supported by National Institutes of Health
Grants DK 42154, DK 37922, and HD 34449 (to W. L. M.).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: Dept. of Pediatrics,
Bldg. MR-IV, Rm. 209, University of California, San Francisco, San
Francisco, CA 94143-0978. Tel.: 415-476-2598; Fax: 415-476-6286.
Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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