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Journal of Lipid Research, Vol 36, 2419-2432, Copyright © 1995 by Lipid Research, Inc.
M Crestani, D Stroup and JY Chiang
The transcriptional regulation of the rat cholesterol 7 alpha- hydroxylase
gene (CYP7) by hormones and signal transduction pathways was studied by
transient transfection assay of the promoter activity. HepG2 cells were
transfected with deletion mutants of the CYP7 upstream region linked to the
luciferase reporter gene. The transcription of CYP7/luciferase chimeric
genes was higher in confluent than in subconfluent cultures of HepG2 cells.
Glucocorticoid receptors, in the presence of dexamethasone, up-regulated
the CYP7 gene through two regions located between -3262 and -2803, and
between -344 and -222, respectively. Thyroid hormones did not have any
effect on the promoter activity. Insulin inhibited the promoter activity
through sequences located between -344 and -222, and abolished the
stimulation by dexamethasone. Hence, the insulin effect was dominant over
that of glucocorticoids. Treatment of transfected HepG2 cells with phorbol
12- myristate 13-acetate (PMA), a known activator of protein kinase C
(PKC), resulted in a time-dependent inhibition of the CYP7 promoter
activity. The negative phorbol ester-response sequences were mapped between
-344 and -222, and between -200 and -161, respectively. The CYP7 promoter
activity was induced nearly 5-fold by all-trans-retinoic acid through
sequences in the region from -200 to -129. Finally, cyclic AMP and protein
kinase A (PKA) stimulated the expression of the CYP7/luciferase gene
through multiple sequences in the distal and proximal regions, and both
positive and negative response regions were mapped. Our results revealed
that the -416 fragment of the rat CYP7 gene confers the activation by
glucocorticoids and retinoic acid, and inhibition by insulin, phorbol
esters and cAMP. It appears that this proximal promoter may contain a
pleiotropic domain that regulates the effects of multiple signals.
ARTICLES
Hormonal regulation of the cholesterol 7 alpha-hydroxylase gene (CYP7)
Department of Biochemistry and Molecular Pathology, Northeastern Ohio Universities College of Medicine, Rootstown 44272-0095, USA.
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