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Downregulation of GATA-1 expression during phorbol myristate acetate-
induced megakaryocytic differentiation of human erythroleukemia cells
W Dai and MJ Murphy
Molecular Biology Laboratory, Hipple Cancer Research Center, Dayton, OH
45439-2092.
Phorbol myristate acetate (PMA) induces the expression of megakaryocyte
and/or platelet proteins during terminal differentiation of human
erythroleukemia (HEL) cells. However, it is not established whether
megakaryocytic differentiation is accompanied by the downregulation of the
major erythroid transcription factor GATA-1 and the concomitant loss of the
erythrocytic phenotype. Studies of the molecular mechanism of PMA-induced
differentiation in HEL cells showed that when HEL cells are treated with
PMA, they dramatically decrease the expression of the erythroid-specific
gene glycophorin A at the mRNA level but apparently not at the steady-state
protein level. In addition, a gel mobility shift assay was used to
demonstrate that GATA-1, a major erythroid transcription factor normally
present at high levels in HEL cells is downregulated after treatment with
PMA. In contrast, the DNA-binding activities of transcription factors AP-1
and SP-1 are upregulated by PMA treatment of HEL cells. Furthermore,
Northern blot analysis shows that PMA also downregulates the steady-state
level of GATA-1 mRNA in HEL cells. The coordinated negative regulation of
glycophorin A mRNA and GATA-1 expression after PMA treatment suggests that
downregulation of GATA-1 expression may be partially responsible for the
loss of the erythroid phenotype during megakaryocytic differentiation. The
reported data also suggest that GATA-1 activity may not be essential for
obtaining megakaryocytic phenotype during terminal differentiation in HEL
cells.
Volume 81,
Issue 5,
pp. 1214-1221,
03/01/1993
Copyright © 1993 by The American Society of Hematology
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