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J. Biol. Chem., Vol. 267, Issue 23, 16719-16726, Aug, 1992
FM Stanley
An insulin response element (IRE) has been identified in the prolactin gene
using chimeric plasmids in which prolactin promoter DNA directs expression
of the bacterial chloramphenicol acetyltransferase gene. A series of
5'-deletion constructs starting between positions -173 and - 106 and
extending through position +75 of the prolactin gene were all stimulated
greater than 10-fold by physiological concentrations of insulin in rat
pituitary tumor GH4 cells. However, insulin did not stimulate constructs
starting at positions -96 and -46, suggesting that the IRE of the prolactin
gene may be located in region -106/-96. Insulin stimulation of
prolactin-chloramphenicol acetyltransferase constructs requires
cotransfection with a human insulin receptor expression vector. Estimation
of insulin receptor levels by beta- subunit phosphorylation indicates that
receptor levels are increased approximately 50-fold following transfection
with the human insulin receptor expression vector. This requirement for
cotransfection suggests that the endogenous receptor levels may not be
adequate to couple the response of transfected genes to insulin. Gel
mobility shift experiments reveal a nuclear factor from GH4 cells that
specifically associates with prolactin DNA fragment -106/-87. The amount or
binding activity of this factor is increased following insulin treatment of
cells. The concordance between functional and binding analyses of the
prolactin promoter confirms the presence of an IRE in region -106/-87. The
insulin-sensitive DNA-binding factor may mediate effects of insulin on
prolactin gene transcription.
An element in the prolactin promoter mediates the stimulatory effect of insulin on transcription of the prolactin gene
Department of Medicine, New York University Medical Center, New York 10016.
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