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(Received for publication, May 13, 1996, and in revised form, August 8, 1996)
From the Previous studies have demonstrated that
insulin-like growth factor-I (IGF-I) increases elastin gene
transcription in aortic smooth muscle cells and that this up-regulation
is accompanied by a loss of protein binding to the proximal promoter.
Sp1 has been identified as one of the factors whose binding is lost,
and in the present study we show that Sp3 binding is also abrogated by
IGF-I, but in a selected manner. In functional analyses using Drosophila SL-2 cells, Sp1 expression can drive
transcription from the elastin proximal promoter, while co-expression
of Sp3 results in a repression of Sp1 activity. Footprint and gel shift analyses position the IGF-I responsive sequences to a putative retinoblastoma control element (RCE). Mutation of the putative RCE
sequence as assessed by transient transfection of smooth muscle cells
results in an increase in reporter activity equal in magnitude to that
conferred by IGF-I on the wild type promoter. Together these results
support the hypothesis that IGF-I-mediated increase in elastin
transcription occurs via a mechanism of derepression involving the
abrogation of a repressor that appears to be Sp3 binding to the
RCE.
Volume 271, Number 46,
Issue of November 15, 1996
pp. 28853-28860
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
A ROLE FOR Sp3 ACTING AS A REPRESSOR OF ELASTIN GENE
TRANSCRIPTION
,
,
,
,
Department of Biochemistry, Boston
University School of Medicine, Boston, Massachusetts 02118 and the
¶ Department of Anatomy and Histology, School of Dental Medicine,
University of Pennsylvania, Philadelphia, Pennsylvania 19104
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