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Forced GATA-1 Expression in the Murine Myeloid Cell Line M1:
Induction of c-Mpl Expression and Megakaryocytic/Erythroid
Differentiation
Yuji Yamaguchi,
Leonard I. Zon,
Steven J. Ackerman,
Masayuki Yamamoto, and
Toshio Suda
From the Department of Cell Differentiation, Institute of Molecular
Embryology and Genetics, Kumamoto University School of Medicine,
Kumamoto, Japan; the Division of Hematology/Oncology, The Children's
Hospital, Harvard Medical School, Boston, MA; the Department of
Biochemistry, University of Illinois at Chicago, Chicago, IL; and the
Center of Tsukuba Advanced Research Alliance, University of Tsukuba,
Tsukuba, Japan.
The "zinc-finger" transcription factor GATA-1 was first shown
in cells of erythroid lineage. It is also expressed in cells of other
hematopoietic lineages including megakaryocytes, mast cells, and
eosinophils. GATA-1 is now considered to be one of the central
regulators in hematopoietic cell differentiation. To further analyze
the role of GATA-1 in controlling differentiation from hematopoietic
stem cells, we investigated the phenotypic changes induced by the
overexpression of murine GATA-1 in the murine myeloid leukemic cell
line, M1. Forced expression of GATA-1 induced the appearance of
erythroid cells and megakaryocytes as assessed by cellular morphology,
acetylcholinesterase activity, and expression of platelet factor 4 and
 -globin mRNA synthesis. Because the c-mpl ligand, thrombopoietin,
plays an important role in megakaryopoiesis, the expression of c-mpl
and c-mpl ligand (thrombopoietin) mRNA was analyzed by Northern blot
and reverse transcription-polymerase chain reaction (RT-PCR) in M1
cells overexpressing GATA-1. The c-mpl ligand mRNA was equally
expressed both in parental M1 cells and in those transfected with the
GATA-1 expression vector. In contrast, the mRNA expression of c-mpl was
increased only in GATA-1 expressing M1 cells differentiated towards
erythroid and megakaryocyte lineages. The increased expression of c-mpl
mRNA induced by GATA-1 raised the question as to whether or not GATA-1 transactivated the c-mpl promoter. The activity of the c-mpl promoter in the presence of cotransfected GATA-1 was significantly increased compared with that of the control. A plasmid with the mutated GATA-binding site did not show transactivation ability in the cotransfection with a GATA expression vector. These findings suggest that the upregulation of c-mpl induced by GATA-1 expression in M1 cells
is closely associated with erythroid and megakaryocytic differentiation.
Blood, Vol. 91 No. 2 (January 15), 1998:
pp. 450-457
© 1998 by The American Society of Hematology.
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