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Blood, Vol. 93 No. 8 (April 15), 1999:
pp. 2637-2644
Regulation of the Megakaryocytic Glycoprotein IX Promoter by the
Oncogenic Ets Transcription Factor Fli-1
L. Scot Bastian,
Boguslaw A. Kwiatkowski,
John Breininger,
Susan Danner, and
Gerald Roth
From the Hematology Section, Medical and Research Services, VA
Puget Sound Health Care System and the Divisions of Hematology
and Oncology, Department of Medicine, University of Washington,
Seattle, WA.
Glycoprotein (GP) IX is a subunit of the von Willebrand receptor,
GPIb-V-IX, which mediates adhesion of platelets to the subendothelium of damaged blood vessels. Previous characterization of the GPIX promoter identified a functional Ets site that, when disrupted, reduced
promoter activity. However, the Ets protein(s) that regulated GPIX
promoter expression was unknown. In this study, transient cotransfection of several GPIX promoter/reporter constructs into 293T
kidney fibroblasts with a Fli-1 expression vector shows that the
oncogenic protein Fli-1 can transactivate the GPIX promoter when an
intact GPIX Ets site is present. In addition, Fli-1 binding of the GPIX
Ets site was identified in antibody supershift experiments in nuclear
extracts derived from hematopoietic human erythroleukemia cells.
Comparative studies showed that Fli-1 was also able to transactivate
the GPIb and, to a lesser extent, the GPIIb promoter. Immunoblot
analysis identified Fli-1 protein in lysates derived from platelets. In
addition, expression of Fli-1 was identified immunohistochemically in
megakaryocytes derived from CD34+ cells treated with the
megakaryocyte differentiation and proliferation factor, thrombopoietin.
These results suggest that Fli-1 is likely to regulate lineage-specific
genes during megakaryocytopoiesis.
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