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The human CD34 hematopoietic stem cell antigen promoter and a 3' enhancer
direct hematopoietic expression in tissue culture
TC Burn, AB Satterthwaite and DG Tenen
Hematology/Oncology Division, Beth Israel Hospital, Boston, MA 02215.
The human CD34 hematopoietic stem cell antigen is a highly glycosylated
type 1 membrane protein of unknown function. CD34 is expressed on 1% to 4%
of bone marrow cells, including pluripotent stem cells and committed
progenitors of each hematopoietic lineage. CD34 has also been shown to be
expressed on the small vessel endothelium of a variety of tissues and on a
subset of bone marrow stromal cells. We have chosen to use the human CD34
gene as model to examine the transcription factors and cis- elements
required for stem cell/progenitor cell-specific gene regulation. We show
here that the CD34 gene is transcriptionally regulated in tissue culture
cells. Using a luciferase reporter gene, we have isolated and characterized
an active CD34 promoter. A CD34- luciferase construct, containing 4.5 kb of
5' flanking DNA from a CD34 genomic clone, was 30-fold more active in CD34+
tissue culture cells than in HeLa cells. Sequences from the 3' end of the
CD34 gene were shown to have enhancing activity in CD34+ T-lymphoblastic
RPMI-8402 cells and not in CD34- U937 cells or in nonhematopoietic HeLa
cells. We also show that a cytidine-guanosine island in the 5' end of the
CD34 gene is heavily methylated in two CD34- hematopoietic cell lines and
demethylated in two CD34+ cell lines. Analysis of the CD34 promoter should
result in the identification of stem cell/progenitor cell- specific
transcription factors and should provide a means to direct the expression
of heterologous genes in hematopoietic stem cells and progenitors.
Volume 80,
Issue 12,
pp. 3051-3059,
12/15/1992
Copyright © 1992 by The American Society of Hematology
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