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J Biol Chem, Vol. 273, Issue 48, 31844-31852, November 27, 1998
From the The human hemopoietic cell kinase (HCK) is a
member of the src family of protein tyrosine kinases
specifically expressed in myeloid cells and to a minor extent in
B-lymphoid cells. HCK expression is up-regulated at the transcriptional
level during myeloid differentiation of hematopoietic cells. To
elucidate the molecular basis of the differential HCK gene
expression, the genomic region containing the HCK promoter
was isolated and functionally characterized. A DNA fragment containing
101 base pairs of the 5'-flanking sequence showed strong promoter
activity in the macrophage cell line RAW264 but was inactive in the
non-monocytic cell lines HUT-78 and NIH-3T3. Site-directed mutagenesis
of the proximal promoter region showed that two GC-rich sequence
elements are essential for transcriptional activity in myeloid cells.
Electrophoretic mobility shift analysis using nuclear extracts obtained
from RAW264 cells and from the promonocytic cell line U-937 revealed
the formation of at least three distinct protein-DNA complexes at each
of these sites, one of which was found to contain the transcription
factor Sp1. Expression of a reporter gene linked to the
The Transcription Factor Sp1 Regulates the Myeloid-specific
Expression of the Human Hematopoietic Cell Kinase (HCK)
Gene through Binding to Two Adjacent GC Boxes within the
HCK Promoter-Proximal Region
,
Laboratory for Molecular Virology,
Georg-Speyer-Haus, D-60596 Frankfurt, Germany and the
§ Paul-Ehrlich-Institut, 63225 Langen, Germany
101
HCK promoter region was up-regulated by Sp1, but not by
other members of the Sp1 family of transcription factors, in
Drosophila Schneider cells. A synergistic effect on
HCK promoter activity was observed at high concentrations of Sp1. Our results show that Sp1 plays an essential role in the regulation of the differential gene expression of the HCK gene.
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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