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Blood, Vol. 92 No. 4 (August 15), 1998:
pp. 1247-1258
Oct-1 Is Involved in the Transcriptional Repression of the von
Willebrand Factor Gene Promoter
Jean-Luc Schwachtgen,
Jacques E. Remacle,
Nathalie Janel,
Reginald Brys,
Danny Huylebroeck,
Dominique Meyer, and
Danièle Kerbiriou-Nabias
From INSERM U143, Unité de Recherches sur l'Hémostase et
la Thrombose, Hôpital de Bicêtre, Bicêtre, France;
and the Laboratory of Molecular Biology (Celgen), University of Leuven
and Department of Cell Growth, Differentiation and Development,
Flanders Interuniversity Institute for Biotechnology, Leuven, Belgium.
The negative regulation of transcription of the human von Willebrand
factor (vWF) gene was investigated in human umbilical vein endothelial
cells (HUVECs) and HeLa cells. A fragment spanning  89 to +244
nucleotides (nt), containing the first exon, is active in HUVECs only
but not in HeLa cells. The activity of this promoter is sharply reduced
by mutagenesis of the GATA binding site at +221. Extension of the
upstream sequences from nt 89 to 142 and to 496 results in
progressive reduction of the activity of the 89 to +244 promoter
identifying a negative regulatory element between nt 142 and 89.
A factor present in nuclear extracts from endothelial and
nonendothelial cells binds to an AT-rich sequence located between nt
133 and 125. Mutagenesis of the AT-rich sequence interferes with
nuclear protein binding and restores the activity of the 142 to
+244 fragment to the level of the 89 to +244 promoter. Binding
of the nuclear protein to the vWF AT-rich sequence in mobility shift
assays is inhibited by competition with a consensus Oct-1 binding site
and with a silencer octamer-like sequence from the vascular cell
adhesion molecule-1 (VCAM-1) promoter. Subsequent supershift
experiments identified Oct-1 as the transcription factor that binds to
vWF and VCAM-1 silencer elements. These results indicate that Oct-1
acts as a transcriptional repressor of promoters of genes expressed in
endothelial cells.
© 1998 by The American Society of Hematology.
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