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(Received for publication, November 12, 1996, and in revised form, March 19, 1997)
From the Production of the placental
hormone, chorionic gonadotropin (CG), increases dramatically as
cytotrophoblasts fuse to form syncytiotrophoblasts. The CG
Volume 272, Number 24,
Issue of June 13, 1997
pp. 15405-15412
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.
- and
-Subunit Promoters by AP-2
,
,
,
and
Division of Endocrinology, Metabolism, and
Molecular Medicine, Northwestern University Medical School, Chicago,
Illinois 60611, the ¶ Department of Endocrinology, Children's
Hospital Medical Center, University of Cincinnati College of Medicine,
Cincinnati, Ohio 45229, and the 
Department of Biology, Yale
University, New Haven, Connecticut 06520
- and
-promoters are both responsive to cAMP, although the kinetics of
cAMP stimulation are different. In an effort to understand the
mechanisms of coordinate induction of these genes, AP-2 binding sites
were identified in the promoter regions of the and CG genes.
AP-2 bound to the upstream regulatory element (
186 to 156 base
pairs (bp)) in the -promoter and to several different regions of the
CG promoter, including footprints 2 and 4B (FP2, 311 to 279 bp;
FP4B, 221 to 200 bp). AP-2 antibodies induced supershifts of these
complexes, confirming the identity of the protein-DNA complex. In JEG-3
cells, which contain abundant AP-2, mutations in these CG AP-2 sites
reduced basal activity and decreased cAMP stimulation. In
AP-2-deficient Hep-G2 cells, co-transfection of AP-2 stimulated
expression of the CG promoter 10-20-fold, and the -promoter was
induced by 3-6-fold. Mutations that eliminate AP-2 binding to CG
FP4B reduced AP-2 stimulation by more than 80%, whereas mutations in
FP2 reduced AP-2 stimulation by less than 50%. Analyses of AP-2
mutants revealed a requirement for the DNA binding/dimerization domain
and the amino-terminal proline-rich and acid-rich transactivation
domains for stimulation of the CG promoter. Primary cultures of
placental cytotrophoblasts were differentiated into
syncytiotrophoblasts in vitro to examine AP-2 expression by
reverse transcriptase-polymerase chain reaction. AP-2 mRNA levels
increased by day 2 and continued to rise in parallel with a marked
increase in and CG gene expression. We conclude that both the
and CG promoters contain binding sites for AP-2 and suggest that
this transcription factor provides a mechanism for coordinating the
induction of these genes during placental cell differentiation.
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