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Human eosinophil Charcot-Leyden crystal protein: cloning and
characterization of a lysophospholipase gene promoter
HI Gomolin, Y Yamaguchi, AV Paulpillai, LA Dvorak, SJ Ackerman and DG Tenen
Department of Medicine, Beth Israel Hospital, Boston, MA 02215.
The Charcot-Leyden crystal (CLC) protein is a lysophospholipase expressed
exclusively by eosinophils and basophils. During eosinophilic
differentiation of eosinophil-committed cell lines, CLC steady state mRNA
levels increase significantly. This increased expression is
transcriptionally regulated during butyrate induction of an eosinophilic
subline (C15) of the promyelocytic leukemia cell line HL- 60, as shown by
nuclear run-on assays. The transcriptional start site of the CLC gene was
identified 43 bp upstream of the 5' end of the longest available cDNA
sequence. The gene encoding CLC protein was cloned from a chromosome
19-specific library and a fragment overlapping the transcriptional start
site was isolated and sequenced. Plasmid constructs (in the pXP2 luciferase
expression vector) containing 411 and 292 bp of genomic sequence upstream
of the CLC transcriptional start site directed reporter gene expression in
transient transfections of HL-60-C15 cells, as well as other myeloid (U937)
and nonmyeloid (HeLa and RPMI 8402) cell lines. However, the differential
expression of the two CLC promoter constructs in these cell lines suggests
that the -292 to -411 bp region of the promoter may confer some specificity
for expression in the eosinophil lineage. The CLC promoter sequence
contains two consensus GATA binding sites, a purine-rich sequence that
presents potential binding sites for PU.1, a member of the ets family of
genes, as well as sequences described in other myeloid-specific promoters.
This is the first demonstration of a functional eosinophil promoter that
could serve as a model for identifying DNA elements and trans-activating
factors that regulate gene expression during the commitment and
differentiation of the eosinophil lineage.
Volume 82,
Issue 6,
pp. 1868-1874,
09/15/1993
Copyright © 1993 by The American Society of Hematology
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