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(Received for publication, January 18, 1996, and in revised form, February 28, 1996)
From the The small leucine-rich proteoglycan biglycan is
involved in several physiological and pathophysiological processes
through the ability of its core protein to interact with other
extracellular matrix molecules and transforming growth factor-
Volume 271, Number 26,
Issue of June 28, 1996
pp. 15787-15795
©1996 by The American Society for Biochemistry and Molecular Biology, Inc.
and
Institute of Anatomy, University of Hamburg,
20246 Hamburg, Federal Republic of Germany and the ¶ Medical
Laboratory Dr. H. R. Ebersold, 3001 Bern, Switzerland
(TGF-). To learn more about the regulation of biglycan core protein
expression, we have cloned and sequenced 1218 base pairs from the
5
-flanking region of the human biglycan gene, demonstrated functional
promoter activity, and investigated the molecular mechanisms through
which various agents modulate its transcriptional activity. Sequencing
revealed the presence of several cis-acting elements including multiple
AP-2 sites and interleukin-6 response elements, a NF-
B site, a
TGF- negative element, and an E-box. The TATA and CAAT box-lacking
promoter possesses many features of a growth-related gene,
e.g. a GC-rich immediate 5 region, many Sp1 sites, and the
use of multiple transcriptional start sites. Transient transfections of
the tumor cell lines MG-63, SK-UT-1, and T47D with various biglycan
5-flanking region-luciferase reporter gene constructs showed that the
proximal 78 base pairs are sufficient for full promoter activity.
Several agents among them interleukin-6, and tumor necrosis factor-
.
were capable of altering biglycan promoter activity. However, in MG-63
cells, TGF-1 failed to increase either activity of the biglycan
promoter constructs or specific transcription from the endogenous
biglycan gene. Since TGF-1 also did not alter the stability of
cytoplasmic biglycan mRNA as determined from Northern analysis
after inhibition of transcription with
5,6-dichloro-1-D-ribofuranosylbenzimidazole, an as yet
unidentified nuclear post-transcriptional mechanism was considered
responsible for the TGF- effect in this cell type. These results
might help to elucidate the molecular pathways leading to pathological
alterations of biglycan expression observed in atherosclerosis,
glomerulonephritis, and fibrosis.
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