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J. Biol. Chem., Vol. 266, Issue 32, 21428-21433, 11, 1991
S Richard and HH Zingg
Retinoids are known to have profound effects on cellular differentiation
and embryo pattern formation. In the adult organism, retinoid acid (RA)
receptors are present in a large variety of tissues, including brain.
However, little is known of the precise roles of RA at these different
sites. In the present study we have identified a novel potential target of
RA action by identifying an RA response element (RARE) in the human
oxytocin (OT) gene promoter. We have used DNA- mediated gene transfer
techniques to introduce various portions of the OT 5'-flanking sequences
next to the chloramphenicol acetyltransferase (CAT) gene in neuroblastoma
cells. RA elicited a marked stimulation of the transcriptional activity of
the OT promoter in cells cotransfected with either the human RA receptor
alpha, beta, or gamma. In cells cotransfected with the RA receptor alpha,
the ED50 of this response was 5 x 10(-10) M. The RA response could also be
conferred to a heterologous promoter independent of orientation.
5'-Deletions as well as site-directed mutations demonstrated that four
TGACC motifs, located at -162, -156, -103, and -83 in the OT promoter, are
necessary for optimal RA induction. Mutation or deletion of any of these
elements reduces significantly the RA response. Interestingly, the first
two TGACC motifs overlap with the estrogen response element that we have
previously characterized in this gene. Furthermore, the TGACC motif located
at -83 overlaps with the CCAAT box. We further demonstrate that in
neuroblastoma cells transfected with an RAR alpha expression vector
expression of the endogenous OT gene is stimulated greater than 4-fold in
response to RA. Our studies constitute the first report of a RARE in a
neuropeptide gene and define a mechanism by which OT gene expression can be
modulated by retinoic acid.
Identification of a retinoic acid response element in the human oxytocin promoter
Laboratory of Molecular Endocrinology, Royal Victoria Hospital, McGill University, Montreal, Quebec, Canada.
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