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Characterization of the human platelet/endothelial cell adhesion molecule-1
promoter: identification of a GATA-2 binding element required for optimal
transcriptional activity
RJ Gumina, NE Kirschbaum, K Piotrowski and PJ Newman
Blood Research Institute, The Blood Center of Southeastern Wisconsin,
Milwaukee 53201-2178, USA.
Platelet/endothelial cell adhesion molecule-1 (PECAM-1) is a 130-kD member
of the Ig gene superfamily that is expressed on platelets, endothelial
cells, and certain leukocyte subsets. To examine the factors controlling
vascular-specific expression of PECAM-1, we cloned the 5'-flanking region
of the PECAM-1 gene and analyzed its transcriptional activity. 5'-Rapid
amplification of cDNA ends (5'-RACE) analysis showed that transcription
initiation occurred at several closely spaced nearby sites originating
approximately 204 bp upstream from the translation start site. Analysis of
the sequence immediately upstream from the transcription initiation site
(TIS) showed no canonical TATA or CAAT elements, however an initiator
element commonly found in TATA-less promoters encompassed the TIS.
5'-serially truncated PECAM-1 promoter segments cloned in front of a
luciferase reporter drove transcription in both a lineage- and
orientation-specific manner. Putative cis-acting control elements present
within a 300-bp core promoter included two ets sites, an Sp1 site, tandem
E-box domains, two GATA-associated sites (CACCC), an AP-2 binding site, and
a GATA element at -24. Mutational analysis showed that optimal
transcriptional activity required the GATA sequence at position -24, and
gel-shift assays further showed that the GATA-2 transcription factor, but
not GATA-1, bound to this region of the PECAM-1 promoter. Understanding the
cis- and transacting factors that regulate the tissue-specific expression
of PECAM-1 should increase our understanding of the mechanisms by which
vascular-specific gene expression is achieved.
Volume 89,
Issue 4,
pp. 1260-1269,
02/15/1997
Copyright © 1997 by The American Society of Hematology
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