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Mechanisms of Transcription in Eosinophils: GATA-1, but not GATA-2,
Transactivates the Promoter of the Eosinophil Granule Major Basic
Protein Gene
Yuji Yamaguchi,
Steven J. Ackerman,
Naoko Minegishi,
Masaki Takiguchi,
Masayuki Yamamoto, and
Toshio Suda
From the Department of Cell Differentiation, Institute of Molecular
Embryology and Genetics and the Department of Molecular Genetics,
Kumamoto University School of Medicine, Kumamoto; the Departments of
Biochemistry and Molecular Biology, College of Medicine, University of
Illinois at Chicago; the Department of Biochemistry, Tohoku University
School of Medicine, Sendai; and the Center of Tsukuba Advanced Research
Alliance, University of Tsukuba, Tsukuba, Japan.
Granule major basic protein (MBP) is expressed exclusively in
eosinophils, basophils, and placental trophoblasts. To identify the
cis-elements and transcription factors involved in regulating MBP expression, we subcloned 3.2 kb of sequence upstream of the exon 9 transcriptional start site (P2 promoter) and serial 5 deletions into
the pXP2 luciferase reporter vector. An 80% decrement in promoter
activity was obtained when MBP sequences between bp  117 to 67
were deleted. To identify transcription factors that bind to and
transactivate through the bp 117 to 67 region, we first compared
the upstream genomic sequences of human and murine MBP; a potential
GATA binding consensus site was conserved in the 50-bp region between
the two genes. To determine which GATA proteins bind this consensus
site, we performed electrophoretic mobility shift assays (EMSAs), which
showed that both GATA-1 and GATA-2 can bind to this consensus site. To
determine the functionality of this site, we tested whether GATA-1 and
GATA-2, either individually or in combination, can transactivate the
MBP promoter in the Jurkat T cell line. Cotransfection with a GATA-1
expression vector produced 20-fold augmentation of MBP promoter
activity, whereas GATA-2 had no activity. In contrast, combined
cotransfection of GATA-1 and GATA-2 decreased the ability of GATA-1 to
transactivate the MBP promoter by approximately 50%. Our results
provide the first evidence for a GATA-1 target gene in eosinophils, a
negative regulatory role for GATA-2 in MBP expression, and possibly
eosinophil gene transcription in general during myelopoiesis.
Blood, Vol. 91 No. 9 (May 1), 1998:
pp. 3447-3458
© 1998 by The American Society of Hematology.
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