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Thyrotropin (TSH) increases 3-hydroxy-3-methylglutaryl coenzyme
A (HMG-CoA) reductase gene transcription in FRTL-5 rat thyroid cells,
and the effect of TSH can be mimicked by cAMP.
Sequence analysis of
the rat reductase promoter has revealed a hitherto unnoticed
cAMP-responsive element (CRE)-like octamer. This octamer is located
between 53 and 60 nucleotides downstream of the sterol regulatory
element 1; its first 6 nucleotides are identical to the consensus
somatostatin CRE, and the entire octamer is identical to the fos CRE. A synthetic oligonucleotide containing the HMG-CoA reductase
CRE-like octamer (RED CRE) formed protein-DNA complexes with nuclear
extracts from FRTL-5 cells, which could be prevented by unlabeled
CRE-containing oligonucleotides whose flanking sequences were otherwise
nonidentical. The complexes were specifically supershifted by anti-CREB
antibodies. FRTL-5 cells transfected with a fusion plasmid carrying the
bacterial chloramphenicol acetyl transferase (CAT) under the control of
the HMG-CoA reductase promoter displayed CAT activity, which was
specifically stimulated by TSH. In contrast, CAT activity in FRTL-5
cells transfected with similar constructs carrying mutations in the
reductase CRE was significantly lower and did not increase after TSH
challenge.
We suggest that the HMG-CoA reductase gene contains a
functional CRE, important for TSH regulation of transcription. The data
presented provide the molecular basis for a novel regulatory mechanism
for HMG-CoA reductase gene expression in rat thyroid cells, which
involves the direct effect of cAMP.
Volume 270,
Number 25,
Issue of June 23, pp. 15231-15236, 1995
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
I. IDENTIFICATION AND CHARACTERIZATION OF A CYCLIC AMP-RESPONSIVE
ELEMENT IN THE RAT REDUCTASE PROMOTER
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