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The tissue-specific transcriptional regulation of the megakaryocytic
glycoprotein IIb gene is controlled by interactions between a repressor and
positive cis-acting elements
MH Prandini, F Martin, D Thevenon and G Uzan
CEA, Laboratoire d'Hematologie, INSERM U217, DBMS, Grenoble, France.
Much information on regulation of the transcription of megakaryocytic genes
stems from studies on the glycoprotein IIb (GPIIb) gene, an early and
specific marker of this lineage. Transcriptional activity is controlled by
the association of positive promoter elements corresponding to binding
sites for the transcription factor GATA-1 and a member of the Ets family.
In the present study, we show that these elements are not directly involved
in the control of cell specificity. In contrast, we identified a sequence
located between -170 and -73 that exhibited a repressor activity based on
an analysis of the transcriptional activity of 5'-deleted GPIIb promoter
fragments transfected in the nonhematopoietic HeLa cells. Further analysis
of this repressor by substitution mutagenesis of the -139/-63 region showed
that bases -120/-116 and -102/-93 were required for full repressor
activity. The repressor is able to interact differentially with GPIIb
promoter elements active in the megakaryocytic HEL, the erythroid K562, the
monocytic U937, or the nonhematopoietic HeLa cell lines, indicating that it
controls GPIIb gene tissue specificity. In addition, direct evidence for
tissue-specific interaction between this repressor and the GPIIb -598/ -406
enhancer was obtained when these elements were set in the context of a
heterologous SV40 promoter. Interestingly, the same repressor element
controlling tissue specificity of the GPIIb gene may also control its
temporal expression during megakaryocyte differentiation, based on recent
evidence obtained by Fong and Santoro (J Biol Chem 269:18441, 1994).
Finally, we found that the -120/-116 GPIIb sequence was part of a consensus
motif shared by promoters of other megakaryocyte-specific genes, suggesting
a common repressor mechanism.
Volume 88,
Issue 6,
pp. 2062-2070,
09/15/1996
Copyright © 1996 by The American Society of Hematology
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